WO1991016675A1 - Automated molecular biology laboratory - Google Patents

Automated molecular biology laboratory Download PDF

Info

Publication number
WO1991016675A1
WO1991016675A1 PCT/US1991/002348 US9102348W WO9116675A1 WO 1991016675 A1 WO1991016675 A1 WO 1991016675A1 US 9102348 W US9102348 W US 9102348W WO 9116675 A1 WO9116675 A1 WO 9116675A1
Authority
WO
WIPO (PCT)
Prior art keywords
liquid
pipette tip
pipette
worksurface
container
Prior art date
Application number
PCT/US1991/002348
Other languages
French (fr)
Inventor
G. Richard Cathcart
Thomas Brennan-Marquez
John A. Bridgham
George S. Golda
Harry A. Guiremand
Marianne Hane
Louis Ben Hoff
Eric Lachenmeier
Melvyn N. Kronick
Douglas H. Keith
Paul Eric Mayrand
Michael L. Metzker
William J. Mordan
Lincoln J. Mcbride
John Shigeura
Chen Hanson Ting
Norman M. Whiteley
Original Assignee
Applied Biosystems, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Applied Biosystems, Inc. filed Critical Applied Biosystems, Inc.
Priority to DE69126690T priority Critical patent/DE69126690T2/en
Priority to EP91908369A priority patent/EP0478753B1/en
Publication of WO1991016675A1 publication Critical patent/WO1991016675A1/en

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0099Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor comprising robots or similar manipulators
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5082Test tubes per se
    • B01L3/50825Closing or opening means, corks, bungs
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0098Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01FMIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F2101/00Mixing characterised by the nature of the mixed materials or by the application field
    • B01F2101/23Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00029Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N2035/00079Evaporation covers for slides
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00178Special arrangements of analysers
    • G01N2035/00237Handling microquantities of analyte, e.g. microvalves, capillary networks
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00346Heating or cooling arrangements
    • G01N2035/00356Holding samples at elevated temperature (incubation)
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00346Heating or cooling arrangements
    • G01N2035/00435Refrigerated reagent storage
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04Details of the conveyor system
    • G01N2035/0401Sample carriers, cuvettes or reaction vessels
    • G01N2035/0403Sample carriers with closing or sealing means
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04Details of the conveyor system
    • G01N2035/0474Details of actuating means for conveyors or pipettes
    • G01N2035/0475Details of actuating means for conveyors or pipettes electric, e.g. stepper motor, solenoid
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04Details of the conveyor system
    • G01N2035/0474Details of actuating means for conveyors or pipettes
    • G01N2035/0491Position sensing, encoding; closed-loop control
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G01N35/1011Control of the position or alignment of the transfer device
    • G01N2035/1013Confirming presence of tip
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G01N35/1016Control of the volume dispensed or introduced
    • G01N2035/102Preventing or detecting loss of fluid by dripping
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027General features of the devices
    • G01N2035/1034Transferring microquantities of liquid
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027General features of the devices
    • G01N2035/1048General features of the devices using the transfer device for another function
    • G01N2035/1058General features of the devices using the transfer device for another function for mixing
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027General features of the devices
    • G01N2035/1048General features of the devices using the transfer device for another function
    • G01N2035/1058General features of the devices using the transfer device for another function for mixing
    • G01N2035/106General features of the devices using the transfer device for another function for mixing by sucking and blowing
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/028Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations having reaction cells in the form of microtitration plates
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1004Cleaning sample transfer devices
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1081Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane
    • G01N35/109Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane with two horizontal degrees of freedom
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10STECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00Chemistry: molecular biology and microbiology
    • Y10S435/809Incubators or racks or holders for culture plates or containers
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00Chemistry: analytical and immunological testing
    • Y10T436/11Automated chemical analysis
    • Y10T436/119163Automated chemical analysis with aspirator of claimed structure
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00Chemistry: analytical and immunological testing
    • Y10T436/25Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/2575Volumetric liquid transfer

Definitions

  • the present invention is related to copending international application "ROBOTIC INTERFACE", number PCT/US90/06000, by Harry A. Guiremand, filed October 16, 1990, which is hereby incorporated by reference.
  • the present invention is in the field of apparatus and methods for performing chemical studies and analyses and has particular application to chemistry protocols involving genetic material from samples of DNA.
  • Aspirating liquid into and dispensing liquid from a pipette can be done several different ways. If a liquid is dispensed into air relatively rapidly, the liquid is dispensed at a regular rate, that is, in an analog fashion. If the same liquid is dispensed relatively slowly, the dispensing rate becomes, at some point, incremental. A droplet forms on the tip, grows, and separates from the tip, then another droplet grows and separates. The size of the droplet depends on such variables as the diameters and the design of the tip and the viscosity and surface tension of the liquid being dispensed. The viscosity and surface tension depend on other variables, among them the liquid material and the temperature.
  • the droplet phenomenon affects aspiration of liquid into a pipette from a container of liquid as well. Liquid is aspirated with a pipette below the surface of liquid in a container, but when the tip is withdrawn, a droplet can form on the tip, and affect the accuracy of the aspiration.
  • the effect of the droplet size on accuracy depends on the volume to be aspirated and the droplet volume.
  • the droplet phenomenon has little effect on accuracy. If, however, the amount to be aspirated or dispensed is in the range of, for example, ten times the volume of a single drop, the droplet phenomenon can be serious indeed, and accuracy may be seriously impaired. In the case of biochemical procedures, the sample size and the volume of material to be aspirated and dispensed is typically very small. If a liquid to be handled is quite viscous, such as genomic DNA for example, the droplet problem assumes larger proportions.
  • liquid is to be dispensed into a container, and the container already contains liquid, the pipette tip can be submerged in the liquid in the container, much in the manner that liquid is typically aspirated, then additional liquid may be dispensed in an analog fashion.
  • a new problem in this procedure is that when the pipette is withdrawn from the liquid in the container, an uncontrolled amount of the liquid can adhere to the outside of the pipette and be carried away when the pipette is moved. Again, if the volume to be aspirated is large compared to the amount that adheres to the pipette, the inaccuracy is small.
  • the amount to be aspirated is small, as is typically the case in biochemical procedures, such as DNA sequencing, the amount that adheres to the outside of the pipette may introduce significant error. Also, the further a tip is immersed in a liquid whether aspirating or dispensing, the more liquid can adhere to the tip, and the greater may be the inaccuracy.
  • a robot for moving a pipette to accomplish liquid transfers from container to container is in some respects a simpler problem than manipulating solid objects.
  • a robot to do pipetting requires three degrees of freedom, while some robot devices require as many as seven.
  • biochemical procedures it is generally necessary to access a large number of different sites, and to do so very accurately.
  • Still another problem encountered in the design of such a system is liquid integrity. Even with rapid movement of robotic components and short and compact site design, the large number of samples and large number of steps for each sample, coupled with time required for such things as heating and cooling, dictates that operations must be done over long periods, such as several hours. Given long processing times and small samples, evaporation can be a serious problem, and can cause significant uncontrolled changes in liquid concentration, introducing error. Moreover, open containers invite problems in cross-contamination. Such contamination can be from carryover with pipette operation and also from evaporation and condensation.
  • the apparatus needs to provide a single pipette tip that can be reused to avoid the clumsy, time-consuming, and error- prone process of frequently discarding a tip and loading a new tip, and the problems of cross-contamination caused by single tip use must be addressed.
  • the apparatus and associated methods of operation also must minimize evaporation and cross-contamination.
  • Such an apparatus needs to be integrated with a control system that allows an operator to easily and quickly set up procedures with different variables, different step sequences, and different samples and reagents.
  • laboratory apparatus based on such a liquid handling system to incorporate further techniques, such as temperature control and a separation station, to be able to fully automate specific chemistry protocols such as for gene detection and DNA sample purification.
  • a liquid-handling instrument to transfer liquid between containers supported on a worksurface.
  • the instrument has a pipette system for aspirating and dispensing liquid and a robotic translation system for moving the tip of the pipette into and out of the containers.
  • the pipette system has a sensing system to sense and communicate proximity of the tip to surfaces on the instrument to the control system.
  • the sensing system has a conductive tip connected to a capacitance sensor. The sensing feature lets the robotic system move the tip with the precision needed for aspirating and dispensing very small volumes of liquid.
  • gaugeblock registered to the worksurface for use in calibrating the control system relative to a precise position on the worksurface.
  • the worksurface also has registration cavities so modular stations may be substituted on the worksurface without losing position integrity, which provides for cleaning and sample setup off-line.
  • the instrument has two syringe pumps connected to the common tip, and the pumps have different capacities, so course and fine aspirations and dispenses may be made with the same tip.
  • the robot in an embodiment is a cartesian device driven by electrical drives with two directions of travel in a horizontal plane over the worksurface and a third at right angles to the surface.
  • the control system has an iconic, user-friendly interface for a user to program steps and enter and edit variable values.
  • the icons are arranged in a manner that more primitive icons are nested in higher-order icons such that higher-order icons can be expanded-in-place to show more program detail without losing relationship with position in a program.
  • a duck-billed closure is disclosed for closing a container to minimize exposure of liquid in the container while allowing easy access by a needle-like device.
  • a liquid-handling instrument according to the invention uses containers with the duck-billed closures to help prevent cross-contamination and evaporation.
  • a container with a duck-billed closure is also disclosed for storing and transporting liquids.
  • An automated laboratory of the present invention for performing chemistry protocols is based on the liquid-handling instrument and has heating and cooling systems to control temperature of samples and reagents during processing.
  • the laboratory has a heated and cooled incubation station with coated container cavities and a latching, sealing lid for sealing container cavities while incubating.
  • the laboratory also has a magnetic station for separating paramagnetic particles from liquids, and the magnetic station has a magnet bar moveable vertically between rows of containers of liquid.
  • a method is also provided to transfer discrete droplets of liquid, and another method is provided to aspirate small volumes of liquid while minimizing tip contamination. Yet another method is provided to mix liquids effeciently with apparatus according to the preferred embodiments. Still another method is provided for validating the placement of elements on a worksurface of the present invention.
  • Fig. 1 is a perspective view of an automated laboratory according to a preferred embodiment of the invention.
  • Fig. 2A is a schematic representation of hardware components of a control system in a preferred embodiment.
  • Fig. 2B is a schematic representation to illustrate hardware and software structure for a control system in a preferred embodiment.
  • Fig. 2C is an example of a partial script list as used in the control system.
  • Fig. 2D is a flow diagram showing the flow of primitives for a specific script command called Dispense.
  • Fig. 3A is a perspective view of a robotic arm assembly for movement in the horizontal plane.
  • Fig. 3B is a perspective view of a robotic arm assembly also for movement in the horizontal plane, but at right angles to the movement of the arm of Fig. 3A.
  • Fig 3C is a perspective view, partially in section of a robotic assembly for vertical movement.
  • Fig. 3D is a perspective view in section of the vertical movement assembly showing additional detail.
  • Fig. 3E is a view of a conductive pipette tip in the preferred embodiment.
  • Fig. 4 A is a plan view of a magnetic station.
  • Fig. 4B is an elevation view in section of the plan view of Fig. 4A with a magnet extended.
  • Fig. 4C is a section view similar to Fig. 4B, but with the magnet retracted.
  • Fig. 4D is a section view of a tube of liquid showing a pipette tip and a helical path used for mixing liquid.
  • Fig. 5A is a view of a computer display showing a high-level icon representing an automated chemistry protocol.
  • Fig. 5B is an expansion-in-place of the icon of Fig. 5A.
  • Fig. 5C is an expansion-in-place of one of the icons of Fig. 5B.
  • Fig. 5D is a further expansion-in-place of an icon of Fig. 5C.
  • Fig. 5E is yet a further expansion of an icon of Fig. 5D.
  • Fig. 6A is a schematic representation of some steps of an example chemistry protocol for the preferred embodiment.
  • Fig. 6B is a representation of further steps of the example protocol of Fig. 6A.
  • Fig. 6C is a representation of further steps of the example protocol of Fig. 6 A.
  • Fig. 6D is a representation of still further steps of the example protocol of Fig. 6A.
  • Fig. 7A is a perspective view of an assembly of a duck-billed closure to a container.
  • Fig. 7B is a section through the assembly of Fig. 7A.
  • Fig. 7C is another section through the assembly of Fig. 7A at right angles to the section of Fig. 7B.
  • Fig. 8 is a section view through an assembly of a duck-billed closure and a container showing a pipette tip inserted through the closure.
  • Fig. 9A shows one step of a method for transferring a droplet of liquid with apparatus according to the preferred embodiment.
  • Fig. 9B shows another step of the method of Fig. 9A.
  • Fig. 9C shows yet another step of the method of Fig. 9A.
  • Fig. 9D shows still another step of the method of Fig. 9A.
  • Fig. 10A shows one step of a method for aspirating liquid using apparatus according to a preferred embodiment.
  • Fig. 10B shows another step of the method of Fig. 10A.
  • Fig. IOC shows yet another step of the method of Fig. 10A.
  • Fig. 10D shows still another step of the method of Fig. 10A.
  • Fig. 11 shows a section through a wash station in a preferred embodiment.
  • Fig. 12 shows a section through a container at an incubation station in a preferred embodiment, with a pipette tip inserted into the container cavity.
  • Fig. 1 is a perspective view of a preferred embodiment of an automated laboratory (AL) 11 for performing chemical processes involved in molecular biology.
  • a computer 13 with a CRT monitor 15, a keyboard 17 and a mouse device 19 is connected to the AL.
  • the computer, CRT display, mouse, and keyboard are hardware components of a control system with an operator interface for programming the AL to perform sequences of activities, for starting and stopping processes and sequences of processes and for entering and altering process variables for specific activities.
  • the computer is a Macintosh II CX computer made by Apple Computer of Cupertino, CA, but other computers may also be used.
  • the AL has a closeable, heated, clamped-lid thermal cycling station 21, an actively cooled enzyme storage station 23, a wash station 25, a reagent storage position 27 for storing and presenting frequently used reagents, a DNA sample stage 28, a wash buffer storage 30, and two magnetic particle wash stations 26 and 29 for manipulating paramagnetic particles in suspension in liquid mixtures.
  • a gauge block 24 for use in calibrating the robotic drives for the apparatus.
  • the various stations are arranged on a worksurface 22. Width DI of the worksurface where all of the stations are arranged is about 50 cm and depth D2 is about 35 cm. The height is about 17 cm.
  • the stations on the worksurface are registered in accurately machined cavities relative to the gauge block so modular stations may be interchanged while maintaining information about the position of containers relative to the worksurface.
  • the magnetic particle wash stations shown are not required for the DNA sequencing protocol included in the description of the preferred embodiment, but are useful for other chemistries and illustrate the flexibility of the apparatus and to provide for ability to do chemistry protocols other than the DNA mentioned above.
  • a projected use of the apparatus of the invention is in purification of biological samples, and the magnetic particle wash stations would be used.
  • a portion of the AL at region 46 is shown cut away to better illustrate the components in the work area.
  • Thermal cycling station 21 has a 96 position array of reaction cavities in 8 columns and 12 rows.
  • the representation in Fig. 1 does not show 96 stations for reasons of detail, and the number 96 is convenient, as it is compatible with the 96 well Microliter plate known and used in the industry.
  • the reaction cavities are machined into an aluminum plate that is electrically resistance heated and also has internal water cooling passages and a thermal sensor for feedback control. Temperature is controlled in the range from 4 degrees C. to 100 degrees C. in the preferred embodiment with 1 degree C. per second rate of change.
  • the reaction cavities are coated with Paralene (TM), a largely chemically inert coating for which materials and process are available from Solid Photography, Inc. of Melville, N.Y.
  • TM Paralene
  • a hinged lid has a polymer undersurface such that, when the lid is closed, the reaction cavities are sealed.
  • Each reaction cavity has a machined detail ring to contact the polymer undersurface to effect sealing (see element 285, Fig. 13).
  • the lid is closable automatically and held closed by a latch in the preferred embodiment. Clamping by the latch is necessary to effect an adequate seal on the seal ring.
  • lid drives such as motor and pneumatic drives are useful, and various kinds of latches may be used, such as mechanical or magnetic. Sealing prevents evaporation, which helps to preserve liquid volume integrity and prevent vapor cross-contamination.
  • Enzyme storage station 23 has three 2 by 8 position arrays for 1.5 mL screw-top tubes, such as available from Sordstadt.
  • the block at station 23 has cooling passages for maintaining temperature of stored enzymes at 4 degrees C. with a tolerance of 1 degree C.
  • a top closure is provided for station 23 with holes in the same array as the 48 tube positions, and the holes are slightly larger in diameter than the pipette tip. The top closure helps to maintain the lower temperature desirable for enzyme storage and holds the tubes in place.
  • Wash station 25 is for washing the pipette tip between liquid transfers to avoid carryover type cross-contamination.
  • the wash station is connected to a waste drain and serves also as a disposal station for liquids that must be expelled from a pipette in a process protocol.
  • Reagent storage position 27 has positions for 1.5 mL screw-top tubes and has no active heating or cooling. The number of positions is optional. Typically 48 positions are provided.
  • DNA sample stage 28 has 96 positions in an 8 by 12 array for tubes containing DNA samples, also with no active heating or cooling.
  • Magnetic particle wash stations 26 and 29 each have a 2 by 12 array for 1.5mL microtubes, and station 26 has active heating and cooling, similar to station 21.
  • Each magnetic station has a three- position vertically moving magnet. The magnets are for manipulating paramagnetic particles used in various protocols to capture specific material from solution.
  • Wash buffer storage station 30 has positions for storage containers for buffer storage. Active heating is provided with temperature sensing and control.
  • a cartesian transport apparatus 31 moves a pipette needle 33 of a system for aspirating liquids from containers at the various stations and dispensing liquids at the same or other stations.
  • the pipette system includes two motor-driven syringe pumps 32 and 34 in the preferred embodiment.
  • Pump 32 is for relatively course transfer, and pump 34 is for transfer of precise amounts of liquids.
  • pump 32 has a larger capacity than pump 34, and the capacity varies depending on the application. For example, pump 32 can vary from 250 microliter capacity for some protocols to 5 mililiters for others, and pump 34 typically has a capacity from 50 to 100 times smaller than pump 34.
  • the two syringes have a common source of diluent.
  • TFE tubing is used from the syringes to the pipette probe tip, with an internal volume of l.lmL.
  • the probe is fitted with a highly polished stainless steel tip that can convey about ⁇ microLiter maximum droplet size.
  • the probe tip in the preferred embodiment is made part of a sensing system for determining when the tip approaches or touches a surface.
  • the tip is conductive, and a wire from a capacitance sensing device is connected to a an electrical contact that contacts the probe tip.
  • a signal is provided to the control system whenever the tip contacts a surface on the AL, and with appropriate circuitry, known in the art, proximity to a surface may also be detected without actually touching.
  • One use of the capacitance sensing tip is to sense the surface of liquids when positioning the tip for liquid transfer operations. By sensing a liquid surface and at the same time keeping track of the height of the tip relative to the worksurface, the liquid level, hence the volume of liquid in a container can be determined. Sensing a liquid surface also provides information as to when and where to aspirate and dispense liquid while minimizing tip contamination.
  • sensing tip Another use for the sensing tip is to examine the physical nature of the working area over which the sensing tip may pass. By passing the tip over the working area at a pre- determined height, at which height the tip will encounter no obstacle if all parts are in their proper place, one can validate the working area. If the tip encounters a surface at any place a surface should not be encountered, it is known that there is a part out of position.
  • the control system can be programmed to provide a warning in any such circumstance.
  • Transport device 31 moves along slot 35 passing over the storage and activity stations.
  • the pipette needle is movable along arm 37 of the transport device in the direction of arrow 39 and the transport is movable along slot 35 in the direction of arrow 41 to position the pipette over any container position at any station.
  • the pipette needle is translatable vertically in the direction of arrow 43 so the transport apparatus is a cartesian XYZ mechanism capable of placing the pipette in any container on the AL work surface.
  • a gauge block 24 in one corner of the work area is used for calibrating the control system as to position of the pipette tip.
  • the gauge block and the active sites on the work area are all pinned to the worksurface with accurate known dimensions.
  • the stations on the worksurface are modular in this fashion, such that a station can be easily and quickly removed and another put in its place, or one kind of station may be substituted for another on the worksurface. Making the stations modular and providing accurate registration to the worksurface allows accurate calibration of the robotic elements to workstation positions at all times.
  • the gauge block has a machined surface for each of the three directions of movement of the cartesian robot, and by approaching and sensing each of the three surfaces in turn with the capacitance sensing probe tip, an accurate home position is communicated to the control system at the start of each protocol in the preferred embodiment.
  • the probe tip can be used in the same way to validate positions of stations and elements on the worksurface. As an example, if a tube at a particular site is wedged out of position in a register opening, such as at too great a height above the worksurface, the probe with capacitance sensing can be used to determine that fact and communicate it to the control, which may then signal for appropriate action.
  • the pipette is for aspirating liquid from any one container and dispensing it into any another container. With the pipette, mixtures of various liquids are made and transported to any other container on the AL.
  • the pipette system also serves to agitate liquids in a container to accomplish mixing, by repeated aspirating and dispensing of the liquid in a container, and in some instances by programmed movement of the tip in concert with aspiration and dispensing.
  • Wash station 25 is for washing the pipette tip to avoid cross-contamination.
  • Computer 13, CRT 15, mouse 19 and keyboard 17 are used with the ROBOTIC INTERFACE referenced earlier, which is a unique iconic program, hereinafter called Popframes, to prepare control sequences and establish specific characteristics for the various activities that make up a complete control sequence, as well as to initiate and terminate specific strings of activities. Entries are also made at the computer to relate specific positions at specific stations on the worksurface with specific samples, such as DNA samples, and with specific reagents that are to be stored at specific sites.
  • the iconic control program is described in further detail in another portion of this specification titled "ROBOTIC INTERFACE".
  • Fig. 2A is a block diagram showing control activities and modules in the preferred embodiment. There are many other control configurations that could be used. Computer 13, keyboard 17, mouse 19, and display 15 are connected together in the usual way, and the computer is connected by communication line 47 to a Motorola 68010 Controller PCB 51 located within the AL chassis represented by dotted enclosure 49.
  • a Motorola 68010 Controller PCB 51 located within the AL chassis represented by dotted enclosure 49.
  • Controller PCB 51 accomplishes high level control functions, such as calculations of robot position and interpretation of communication from the computer, and translation of the computer communication into more fundamental control signals for other control hardware.
  • the controller PCB communicates by path 53 with Function PCB 55.
  • the function PCB accomplishes, among other functions, all of the Input/Output (I/O) operations necessary in the control operations.
  • I/O Input/Output
  • sensors on the AL to sense positions of the robot arm, such as mechanical switches.
  • the Function PCB monitors the status of position sensors as digital data and converts that data to computer level signals for the computer part of the control system.
  • the Function PCB monitors analog data communicated by analog sensors on the AL, such as temperature monitoring sensors.
  • the Function PCB converts the analog data to data suitable for the computer portion of the control system.
  • the Function PCB handles all analog-to-digital (A/D) conversion and digital-to-analog (D/A) conversion between the computer portion and actuators and other equipment on the AL.
  • Function PCB 55 communicates along path 57 with the X-Y-Z robot 59, the station modules 61 on the worksurface, the syringe pumps 63 and the capacitance sensor probe 65, and also with Power Driver PCB 67 through path 69.
  • Communications along path 57 are primarily sensor data sent to the Function PCB.
  • Signals along path 69 are primarily signals from the Function PCB to the Power Driver PCB to actuate motions on the AL.
  • An AC Input and Power Supply chassis 54 in the AL receives primary AC power from outside the AL, and has the purpose of dividing, conditioning, and providing power to all the power requirements on the AL, which it does by virtue of on-board power supplies connected to the Power Driver PCB along path 56.
  • the Power Driver PCB has the primary function in the preferred embodiment of switching power to various drivers on the PCB as required for operation, such as to the DC motors that operate the X, Y, and Z motions of the robot.
  • the power to the various parts of the AL is provided primarily along path 58.
  • Fig. 2B is a largely schematic drawing to illustrate in greater detail how communication passes from the computer, a Macintosh II CX in the preferred embodiment, to other control hardware, and to illustrate in more detail the structure of software for accomplishing the tasks.
  • the interface for setup of the AL regarding constants and variable values, and for programming protocols is an iconic program called Popframes.
  • a high level sequence of more basic steps is indicated on monitor 301 of the control computer by an icon such as icon 303.
  • the lines 305 and 307 extending from the icon indicate sequential connection to other icons in a programmed protocol, although other such icons are not shown in Fig. 2B.
  • Each icon developed for Popframes is associated with a command list called a script, and the script for icon 303 is represented in Fig. 2B by enclosure 311.
  • the script for icon is called in the Macintosh hardware.
  • the script is sent to the Motorola 68010 PCB in the AL chassis in the preferred embodiment.
  • Fig. 20 is a short excerpt from a script list.
  • Script is programming protocol available from Apple Computer of Cupertino, CA. and used with Apple computer hardware.
  • the script sent to the Motorola 68010 microprocessor in the preferred embodiment is interpreted there into Forth protocols that are themselves lists of more primitive functions for the AL.
  • For each script step there is a Forth kernel programmed on the 68010, and kernel list 313 shows a selected few of the kernels.
  • Each script step activates a Forth kernel, and a series of primitives is performed in an order often determined by setting of flags and other variables. Communication from the Forth kernels to discrete actuators on the AL is not shown in Fig. 2B.
  • Forth is a well known language often used in the art to program controls for robotic devices, and there are many reference books in the art explaining the structure and use of Forth.
  • Fig. 2D is a flow diagram showing a sequence of more primitive functions associated with one script step called Dispense, which controls dispensing of liquid from the pipette tip.
  • Element 315 the Dispense Script command is the start of the sequence, and there are several decision points, based upon flags that can be set. One such is decision point 317, asking if the KissOff flag is set. If the flag is set, the procedure follows one path, and if not, another path is followed.
  • 'descendSpeed' for example is a rate of travel for the system to use to move the pipette tip downward toward a liquid surface.
  • 'descendSpeed' is a rate of travel for the system to use to move the pipette tip downward toward a liquid surface.
  • many of the primitives are themselves combinations of even more basic functions.
  • element 319, "move down to 'dispenseLevel' at full speed is composed of a sequence that starts the vertical drive, ramps it up to full speed (pre-programmed), ramps it down near the 'dispenseLevel', and stops the drive with the pipette tip at 'dispenseLevel'.
  • Fig. 3A is a perspective view of mechanisms for driving cartesian robot 31 in the X-direction, which is the direction of arrow 41 in Fig. 1.
  • the view of Fig. 3A has the Y-direction and Z- direction mechanisms removed, so the X-direction mechanisms may be better illustrated.
  • X-direction motion is provided by a D.C motor 119 that drives a flexible gear belt 121.
  • a cast frame 123 supports the X-direction drive assembly, and the frame is mounted by conventional fasteners to baseplate 125, which is the baseplate to which stations on the worksurface in Fig. 1 are mounted.
  • the frame is positioned precisely on the baseplate by locator pins, such as pins 127 and 129.
  • Motor 119 is mounted to frame 123, and a pully 131 on the motor shaft drives an intermediate toothed gear belt 133 which in turn drives another pulley 135.
  • Pully 135 is mounted on a shaft through frame 123 in bearings (not shown) and drives yet another pulley 137.
  • Gear belt 121 extends between driven pulley 137 and an idler pulley 139 at a distance greater than the maximum X- direction movement, which is about 45 cm. in the preferred embodiment.
  • a travelling cast carriage 141 is mounted below gear belt 121 on linear bearings arranged such that the carriage rides on a linear guide bar 145, which is fastened also to frame 123.
  • Carriage 141 is attached to one side of gear belt 121 by a clamp 147 such that, as motor 119 causes belt 121 to traverse, the carriage is caused to traverse along bar 145 in the X-direction.
  • Extension 149 from carriage 141 carries optical sensors 151 for sensing flags (not shown) fastened to the AL frame to signal position to the AL control system.
  • the linear bearings are precision bearrings such that the maximum runout from end-to- end does not exceed about .005 inches (.013 cm).
  • reaction cavities for example, at station 21 (Fig. 1) are on about 1 cm centers, and the diameter of each cavitity at the base is about .12 cm.
  • the shaft encoders and bearings used for the X-drive, along with the control system, provide resolution of the robot in the X-direction of .020 mm.
  • Lands 153, 155, 157 and 159 on carriage 141 are machined at a constant height to mount mechanism for Y-direction translation.
  • Fig. 3B shows the Y-direction mechanisms.
  • base plate 161 is the frame for mounting other components, and plate 161 mounts to carriage 141 of Fig. 3A and travels with that carriage.
  • Surface 163 mounts to land 153 of Fig. 3A and surface 165 mounts to land 157 of Fig. 3A.
  • the surfaces that mount to lands 155 and 159 on carriage 141 are not seen in Fig. 3B.
  • Mounting plate 161 is shown as a flat plate for simplicicty, but is typically a casting with reinforcement ribs and the like in the preferred embodiment.
  • Y-drive motion is provided by a D.C. drive motor 167 mounted to a stand 169, that is fastened to plate 161.
  • the motor drives a pulley (not shown) on the motor shaft, which drives a gear belt 171 around an idler pulley 173 rotatably mounted to a standoff near the end of plate 161 opposite the end where the drive motor is mounted.
  • a moving carriage 177 is mounted on linear bearing 179 and constrained to guide along a guide bar 181 affixed to plate 161. Carriage 177 is fastened to belt 171 by a clamp (not shown) similar to clamp 147 of Fig. 3A, such that as motor 167 turns and belt 171 is driven, carriage 177 moves along guide bar 181 in the Y-direction.
  • a Z-direction mechanism 183 is mounted to a guide bar 185 and constrained to guide in a linear bearing 187 mounted to carriage 177 to provide motion in the vertical, or Z-direction.
  • the Z-direction mechanism is driven by a D.C. motor 189 mounted to carriage 177 and turning a pinion 191 which in turn drives a rack 193 that is fastened to the Z-direction mechanism.
  • the Z-direction mechanism protrudes through a slot 195 in plate 161.
  • Fig. 3C shows additional detail of the Z-direction mechanism.
  • Block 197 of Z-drive mechanism 183 serves as a frame for other components.
  • Rack 193 and the guide bar for the vertical guide linear bearing mechanism are attached to block 197.
  • a probe assembly 199 with an outer body 213 is slidably engaged in a multi- diameter cylindrical bore 201 of the body with clearance for a coil spring 203.
  • the bore diameter is smaller at regions 205 and 207, such that the clearance between the outside of body 213 and the guide diameters of the bore is about .1 mm., while the clearance in the region for the coil spring is about 1mm.
  • Spring 203 is captured between a shoulder 209 in block 213 and a shoulder 211 on body 213.
  • Body 213 is limited in vertical travel by shoulder 215 in block 197 and shoulder 211 on body 213.
  • the vertical travel against the spring is for sensing contact with a resisting surface without damaging probe tip 33.
  • Block 197 and body 213 in the preferred embodiment are made of an engineering plastic material to be non-conductive, such as nylon.
  • Tip 33 is stainless steel and brazed in the preferred embodiment to a stainless steel cylinder 217 which fits in a bore in body 213.
  • a stainless steel thumb nut 219 threads onto body 213 and captures cylinder 217.
  • a probe contact 221 connected to wire 223 supplies electrical potential to the probe tip, and is captured between thumb nut 219 and a thumb screw 225.
  • Non-conductive polymer tubing 227 leads from the probe tip to the syringe pumps.
  • Fig. 3D is a vertical section view of the probe assembly shown without the coil spring, contact 221 and the electrical wire.
  • Body 213 in vertical section is shown engaged in block 197 with stainless steel cylinder 217 captured in a bore in body 213 by thumb nut 219 which engages body 213 by threads 229.
  • Thumb screw 225 is shown threaded into thumb nut 219 by threads 231.
  • Ferrule 233 is a separate piece for establishing a seal between the probe tip assembly and the delivery tubing by virtue of pressure applied with the thumb nut.
  • an optical sensor 235 is shown that senses movement of body 213 in block 197.
  • Probe tip 33 is part of a brazed assembly including stainless steel cylinder 217.
  • Overall length D7 in the preferred embodiment is about 87 mm and the length D6 of cylinder 217 is about 20 mm.
  • the diameter D5 of cylinder 217 is about 6.4 mm (.25 inches).
  • the tube portion is made of type 304 stainless steel tubing of about 1.27 mm (.050 inch) outside diameter and about .8 mm (.032 inch) inside diameter.
  • the tube is narrowed so the inside diameter D4 is about .3 mm (.012 inches). Having the diameter at the small dimension for only the tip end length is an advantage in that the flow resistance of the entire tube length is unaffected.
  • the resolution in the X-direction is about .020mm, in the Y-direction about .025 mm, and in the Z- direction about .015 mm.
  • the control system in the preferred embodiment also provides speed ramping that can be varied by an operator through the unique operator interface, and capability to program special motions, such as a helical motion in the Z- direction to facilitate mixing operations.
  • special motions are implemented as combinations of two or more of the basic X, Y, and Z motions.
  • the cartesian robot has a home position in the back, left corner of the work area (facing the AL), with the vertical drive at the full up position.
  • This home position is determined by optical sensors built into each of the three direction mechanisms.
  • a home position protocol is programmed in which the tip is moved slowly to touch each of three reference surfaces on a gauge block (block 24 in Fig. 1), and the robot position is recorded for each of the three points at the time that that capacitance sensing tip touches each of the three reference surfaces. This protocol is performed typically each time a new chemistry protocol is commenced.
  • solutions to be separated are transferred to vials at one of the magnetic wash stations 26 or 29 (Fig. 1).
  • the use of one or the other depends on whether heating or cooling during separation and washing is known to facilitate the process.
  • Precoated particles suspended in a buffer solution are aspirated from a position at one of the reagent storage stations and dispensed into the solutions to be processed at the magnetic wash station.
  • Fig. 4A is a plan view of wash station 26, with heating and cooling capability. There are two rows of twelve tube positions each at the station. In the space between the rows of tubes there is a magnetic bar 237.
  • Fig. 4B shows a section through the station of Fig. 4A taken along section line 4B-4B. Magnetic bar 237 is attached by connector 239 through a screw mechanism (not shown) to a D.C. motor 241. The motor is driven by the control system to move the magnet vertically between the rows of tubes, in the direction of arrow 243.
  • the magnets used are composed of rare earth materials, for example, an alloy of Niobium and Boron with iron, to obtain a high strength magnetic field.
  • the field strength in the area of the inside of the tubes is about 35 million gauss-oersteds.
  • the magnets are raised after the paramagnetic particle suspension is added, and the particles are attracted into closely packed regions that are eventually located near the bottom of the tubes as shown by regions 245 and 247.
  • the ability to move the magnetic bar for the full height of solution in the tubes and to stop it at various points allows the entire solution volume to be swept by the intense field and the particles to be collected into a small area efficiently. It is also advantageous to use a long tube with a small diameter as opposed to a shorter tube with a larger diameter, because the paramagnetic particles have a shorter distance to travel through liquid to be collected. By slowly lowering the magnetic bar the collected particles are moved to the bottom of the tube.
  • wash buffer is aspirated at station 30 and dispensed into each of the tubes at the magnetic wash station where separation is being done.
  • the wash buffer is added with a programmed helical motion from near the bottom of a tube until all of the buffer is added, imparting a stirring action as the buffer is added.
  • Fig. 4D shows a vertical section of one of the tubes of Fig. 4C and the pipette tip of the AL.
  • the helical motion of the tip while dispensing wash buffer is approximated by path 249, and is pre-programmed using motions in all three directions X, Y, and Z.
  • the magnetic bar is raised again to re-collect the particles. The action can be repeated as often as necessary, and is typically done four or five times.
  • a unique program is run on the computer in the preferred embodiment to create control programs, enter and edit variable values, and to initiate and terminate process sequences.
  • the program hereinafter called Popframes, is an iconic program that employs graphic symbols called icons to represent processes, process steps, and other activities, and is described in copending patent application entitled ROBOTIC INTERFACE, Serial No. 07/423,785 referenced earlier.
  • Popframes provides a unique user interface that is useful for handling hierarchical information and for controlling many kinds of process machines and equipment.
  • Popframes has a set of routines allowing a user to select icons representing various activities and to organize the icons into flow schematics representing process flow, with the icons connected on the display with lines.
  • the icons may also be nested such that a relatively complex sequence of activities may be represented by a single icon, and the single icon may be expanded in place to show a connected sequence of icons representing steps in the more complex sequence.
  • the second level icons may also consist of sequences of other icons, also expandable in place, until, at the lowest level, icons represent fundamental process steps.
  • the fundamental steps in the preferred embodiment are typically themselves sequences of even more basic activities. For example, a fundamental step may be a direction by the program to the AL to send the robot arm to a specific position at the DNA stage, station 28 in Fig. 1.
  • the command from the computer to the electronics interface is equivalent to "Go to position X at station 28."
  • the position is a known site to the control system, and sensors tell the control system where the robot arm is before the move.
  • Quick calculation determines the magnitude of the X, Y and Z moves to reach the destination from the starting point.
  • the system then accomplishes the necessary drive sequence with default acceleration and velocity.
  • Fig. 5A shows a screen display 69 in Popframes with a program icon 71 for the Taq DNA sequencing protocol.
  • the single icon represents all of the steps and procedures of the protocol of sequencing DNA templates by the Taq procedure described above.
  • a screen cursor 73 is movable over the area of the screen by moving mouse device 19 over a surface. This is a phenomenon very familiar to those skilled in computer arts.
  • Fig 5B shows the result of expanding the Taq icon in place.
  • the Taq program icon is represented in the expansion by a box 91 surrounding the eight icons shown in sequence. There is a hierarchical relationship between the original icon, which is at the top of the hierarchy, and the sequence of eight icons of Fig. 5B, which are at one level below the top level icon.
  • a user can reverse the expansion process, collapsing a sequence of icons into a higher level icon.
  • the method is by clicking on the close box 93 at the upper left corner of the Taq box within which the eight icons appear. Clicking means that the cursor is moved to the close box, and the mouse button is pressed once.
  • the expansion then collapses back to the original Taq icon at the highest level.
  • the highest level icon does not have a close box, because none is needed, but boxes at all levels below the highest level do have close boxes.
  • Fig. 5C shows the expansion result initiated by double clicking on the Incubate icon in Fig. 5B.
  • the Incubate process is seen to be composed of two distinct steps, step 95 to cycle 10 minutes at 70 degrees C, and step 97, which cycles the temperature after 10 minutes at 70 degrees C. to 10 degrees C.
  • the Incubate icon has become the surround box 103 with a close box 101. The heirarchical relationship of the entire program is still preserved.
  • step 95 by double clicking illustrates yet another feature of the iconic progam in the preferred embodiment.
  • Fig. 5D shows the expansion of step 95 as a variable-entry box 105.
  • Box 105 is at the lowest level of the heirarchical relationships in the iconic scheme, and provides several text fields for entering information for the computer to follow when performing the step.
  • Rack entry field 107 allows a user to enter the name of the rack where the temperature cycling is to be done.
  • a user makes an entry by clicking on the text field, which enables the field for entry, then entering the designation of the rack from the keyboard.
  • the entry field while entry is being made, works much like a word processor. If a mistake is made, the backspace key allows the user to correct the error.
  • Temperature field 109 is for setting the temperature for the temperature step.
  • Ramp field 111 is for setting a ramp rate for changing the temperature.
  • Hold field 113 is for entering a time for holding the temperature at the set temperature.
  • Failsafe field 115 is for entering a temperature range for deviation from the set variables without aborting the process.
  • the expansion has become too broad to be shown on the screen, and the Taq surround box shows terminated at the right edge of the screen.
  • the cursor By placing the cursor inside the Taq surround box, holding down the mouse button and moving the mouse, a user can move the display to show the hidden portion at the right. This is a process called panning in the art. By panning a user can still see all of an expanded program, so information about the heirarchical relationships of the program is always preserved.
  • variable-entry box At the lowest level of expansion of each of the other icons there is a variable-entry box. For example, at the lowest expansion level of the setup box, there is a variable-entry box with fields for the user to relate specific sites at each station to specific samples and reagents that are to be loaded for the analytical sequence.
  • Fig. 5E shows Fig. 5D with programming function menu bar 117 displayed.
  • Figures 6A, B, C and D illustrate a typical biochemical procedure performed on the AL in the preferred embodiment, and is illustrated both as an example of use and as a basis for further description of apparatus and methods in preferred embodiments of the invention.
  • the example illustrated is a proprietary Applied Biosystems, Inc., protocol based on the Sanger termination method for DNA sequencing with Taq polymerase, performed on one single- stranded DNA template.
  • Each column in Fig. 6A, 6B, 6C and 6D represents one step in an automated protocol, with the progression of steps numbered at the top of the columns, reading from the left to the right through the four figures.
  • the liquid volume dispensed to a container in any operation is listed to the right of the container, and the total liquid volume in the container is in parentheses.
  • the protocol involves only three mechanical functions in the automated system: robotic positioning of the pipette tip, small-volume liquid handling through and with the pipette tip, and heating and cooling.
  • the user begins the chemistry by loading tubes of the DNA template to be sequenced and the necessary reagents in the robotic system.
  • the DNA sample tubes are loaded to station 28 (Fig. 1), the DNA stage in the preferred embodiment.
  • station 28 Fig. 1
  • several different DNA templates would be sequenced, and the 96 position array at the DNA stage in the preferred embodiment allows 24 different templates to be sequenced at the same time.
  • step 1 dye-labeled primers are annealed to the DNA template.
  • DNA template is moved from the DNA stage to containers at the thermal cycling station 23 (Fig. 1).
  • One template is prepared for each of the four base types A, C, G and T.
  • Taq sequencing buffer is moved in step 2 from reagent storage station 27 to the containers at the thermal cycling station in the amounts shown in the figure.
  • step 3 the dye-labeled primers are added, and in step 4 pure water is added to each reaction container.
  • step 5 the lid is closed at station 21, heat is applied, and the dye-labeled primers are annealed to the DNA templates at 55 degrees C. for 5 minutes.
  • step 6 the reaction containers are cooled at 20 degrees C. for 20 minutes.
  • steps 7-14 the Taq DNA polymerase synthesizes complimentary DNA chains along the DNA templates to the dideoxynucleotide terminations.
  • the Taq enzyme is deactivated in the alcohol precipitation of steps 15-18.
  • step 18 the product is ready for flourescent sequencing by gel electrophoresis.
  • the robot makes 751 moves, or 31 moves per template.
  • the number of necessary moves provides motivation for automating the protocol.
  • the magnetic wash stations are not needed, but they are useful in other protocols, such as gene scanning.
  • Fig. 7A is a perspective view of a tube closure 253 used in the invention to prevent evaporation of materials during processing, and to provide other advantages.
  • Closure 253 is called a duck ⁇ billed closure, and is shown assembled to a tube 255 of a sort often used for samples, enzymes and reagents.
  • Closure 253 is molded from a flexible material, typically butyl rubber in the preferred embodiment.
  • Such duck-billed closures are a feature useful in many, but not necessarily all, applications of the present invention. The closures are most useful in embodiments where problems related to evaporation are potentially more serious than in other applications.
  • Fig. 7B is a vertical section of the tube and closure shown in Fig. 7A along the section line 7B-7B.
  • Fig. 7C is a vertical section of the same assembly along the section line 7C-7C, taken at a right angle to section 7B-7B.
  • the duck-billed closure in the preferred embodiment has a seal portion 252 with a cavity, usually circular, for enclosing the upper rim of a container to be closed.
  • a flexible duck ⁇ billed portion 254 extending into the container from above, such that a needle-like device, such as the probe tip in the preferred embodiment, may be easily inserted from above and withdrawn to access liquid in the container.
  • the duck-billed closure remains urged against the tip with a bare minimum of opening for possible escape of liquid or vapor.
  • the duck-billed closure closes, and effectively prevents liquid or vapor escape.
  • the outside diameter D8 of the closure in the illustrated embodiment is about 13.2 mm.
  • Dimension D9, the width of the duckbill portion is about 4.8mm.
  • the height D10 of the duckbill closure is about 7.1 mm.
  • the included angle Al of the duckbill portion is about 45 degrees.
  • the wall thickness Dll of the duckbill portion is about ,25mm.
  • Fig. 8 shows the tube and closure of Fig. 7A, B, and C with a probe tip 33 inserted.
  • the tip can penetrate the closure from above with little effort and be withdrawn with little effort as well.
  • penetration or withdrawal there is no mechanism or motion involved more than is involved if there is no closure at all, and the duckbill is caused to open only the exact amount needed to admit the probe.
  • additional mechanism and robotic control must be provided to open and close the lid for access to the contents of a tube.
  • the duckbill closure effectively prevents evaporation, eliminating inaccuracies and cross-contamination that evaporation can cause.
  • the duckbill closure At stations where heat is applied the duckbill closure not only prevents evaporation, but effectively seals against small buildup of pressure inside the tube.
  • reagents and other materials used in the AL can be packaged for transport with the duckbill closure in place, avoiding need to transfer the contents from one container to another during setup of the AL for a protocol. This is useful because it is very common to use a device like the AL at one site and to prepare samples and other materials at another. Moreover, most reagents are prepared by supply houses and sold to laboratories, who seldom prepare their own. The use of a duckbill closure in the original packaging can avoid potential for error and contamination. In the process of packaging with a duckbill closure, a secondary secure cap can be applied for shipment and removed at the use site without disturbing the contents.
  • duckbill closure Another advantage of the duckbill closure is that tubes can be removed from the AL after chemistry protocol and transferred directly to a centrifuge in those cases where centrifuging is desirable.
  • Fig 9A shows the pipette tip 33 in the preferred embodiment with a droplet 257 of liquid formed on the end.
  • the tip and droplet are shown positioned over a vial 259 containing a liquid having a surface 261.
  • a droplet is typically formed by aspirating liquid with the pipette, then driving a syringe pump to dispense just enough liquid to cause a droplet to form.
  • the size of the droplet is determined by such factors as the diameters and material of the pipette tip, the angle, if any, on which the tip is cut, the material to be pipetted, the volume driven by the syringe pump, the temperature, and other factors.
  • Fig. 9B shows the pipette of Fig. 9A lowered toward vial 259 to the point that droplet 257 just touches liquid surface 261 in the vial.
  • Fig. 9C shows the situation a fraction of a second after the droplet touches the liquid surface.
  • the droplet is merging with the liquid in the vial and is still adherent to the tip by virtue of the surface tension of the liquid.
  • Fig. 9D shows the situation after raising the tip.
  • the liquid surface has separated from the liquid still in the pipette tip and from the pipette tip, leaving only a small miniscus 263 at the end of the tip.
  • the droplet conveyance technique is used in the preferred embodiment to transfer discrete volumes of liquid as small as 1 micro-liter.
  • the kiss-off technique is a series of movements for the AL that are programmed into a reusable sequence with an icon, and can be placed in new sequences as required using the Popframes programming interface described above.
  • Another liquid handling technique that has been developed in the preferred embodiment is a technique of accurately aspirating liquids with the pipette tip while minimizing contamination of the tip.
  • the technique is particularly applicable to handling viscous liquids, which are generally more troublesome in liquid handling than are less viscous liquids.
  • Figs. 10 A, B, C, and D show the steps used in this technique.
  • probe tip 33 is positioned over the surface 265 of a liquid to be aspirated, as shown in Fig. 10A.
  • the tip is lowered to touch the surface, sensed by the capacitance sensing ability associated with the probe tip, as shown in Fig. 10B.
  • Aspiration of a programmed amount is accomplished slowly, typically at about 1 micro-liter per second, while the tip is at the surface as shown in Fig. 10B.
  • the rate of aspiration is set to suit the viscosity of the liquid to be aspirated. If the amount to be aspirated is quite small relative to the volume in the container, then the tip position will not have to be adjusted vertically during the aspiration.
  • the position of the tip can be adjusted downward during the aspiration to maintain the relationship of the tip to the liquid surface.
  • the pipette tip can be lowered a fixed small amount to penetrate the liquid surface a minimal amount before aspiration begins.
  • the probe tip is slowly withdrawn, typically at a rate of about 1.5 mm per second. As the tip is withdrawn, initially liquid still clings to the tip as shown in Fig. IOC, and this condition varies depending on the viscosity and surface tension of the liquid. The tip and the liquid separate as withdrawal continues, as shown in Fig. 10D. The probe is then moved to wherever is required to dispense the liquid that has been aspirated.
  • the aspiration technique is a series of movements for the AL that are programmed into a reusable sequence with an icon, and can be placed in new sequences as required using the Popframes programming interface described above.
  • wash station 25 is used as needed between liquid transfers to cleanse the tip before a different reagent or sample material is transferred.
  • the tip can be washed both inside and outside.
  • Fig. 11 shows the pipette tip in position to wash the tip at wash station 25.
  • the wash station includes a body 279 with a fountain 271, a well 273 and a drain 275. Body 279 is shown in section so the position and nature of other components may be seen.
  • the fountain is a generally cylindrical bore of a depth and diameter such that wash buffer dispensed from the pipette tip will backflow and wash the outside of the tip.
  • the tip dimensions may vary for different protocols and purposes.
  • the tip is about .6 mm in outside diameter for a length from the end of about 6 mm.
  • the depth of the fountain D12 is about 6 mm and the diameter D13 about 1.2 mm.
  • the requirement is to provide an annulus for liquid backflow around the outside of the tip to backwash the outside of the tip beyond the length that will be inserted into a liquid on the AL.
  • Wash buffer dispensed from the pipette tip at the wash station to cleanse the tip backflows vertically in annulus 277 and spills over into well 273, where it drains through drain 275 to a waste container below the worksurface in the AL.
  • the wash station serves also as a waste disposal station. For waste disposal from the pipette tip without washing the tip, the tip is positioned over well 273 and the waste is dispensed to drain 275. For waste disposal it is not needed to position the tip in the fountain.
  • Fig. 12 is a section through one of the reaction vessels 281 at incubation station 21 with the pipette tip shown inserted into the vessel.
  • the reaction vessel is machined with sloping sides such as side 283, a raised lip 285, and a cylindrical chamber 287.
  • the material for the plate is aluminum, for the desirable heat transfer characteristics, and the surface is coated with Paralene (TM) before use so the aluminum cannot react with the materials placed in the reaction volume.
  • TM Paralene
  • the raised lip is so the lid, which has a sheet of flexible material on the undersurface, butyl in the preferred embodiment, will seal to the reaction vessel when the lid is closed.
  • Chamber 287 is where material is actually deposited and where reaction is accomplished.
  • the plate at station 21 into which the reaction vessels are machined is a replacable modular unit, so plates can be assembled to the AL with reaction vessels of different sizes for different purposes.
  • the vessel shown in Fig. 12 is for reaction volumes of about 50 micro-liters. Diameter D14 is about 1.25 mm and depth D15 is about 1.52 mm. Total depth D16 is about 8 mm, diameter D17 is about 6 mm, and angle A2 is about 40 degrees in the embodiment shown.
  • Southern blotting a very widely practiced technique in the molecular biology laboratory, is used to determine the length of DNA fragments homologous to a particular DNA probe (3). It has proven extremely valuable in tracking genetic diseases and identifying the presence of specific forms of genes in complex samples such as human genomic DNA (4,5).
  • the specific chemical steps required for Southern blotting (such as blotting transfer to membranes, membrane handling, and autoradiography) are not amenable to automation, though some attempts have been made (6).
  • a novel chemistry has been developed which produces results equivalent to those from a Southern blot experiment; the process is solution based which allows for total automation by a liquid handling robot. The details of this chemistry are described elsewhere (7,8).
  • the new liquid-based methodology involves the following steps: 1) genomic DNA is simultaneously digested with a restriction enzyme and fluorescently labeled; 2) the genomic DNA is denatured and a biotin labeled probe is hybridized in solution to specific target molecules within the population of restricted genomic DNA fragments; 3) the specific hybrids are captured onto the surface of streptavidin functionalized paramagnetic particles while the remainder of the restricted genomic DNA population is not; 40 non-specific genomic DNA molecules in solution and bound to the particles are removed by stringent washing; 5) the captured hybrid/paramagnetic particle complexes are loaded directly into the well of a denaturing electrophoresis gel and the released labeled target molecules are detected when they electrophoreses past a laser scanned region a defined distance from the sample loading well; 6) collected fluorescent light is measured and the resultant data is analyzed.
  • Other workers have described techniques where hybridization precedes electrophoresis but these techniques did not
  • the chemical methodology described above lends itself to automation with a robotic liquid handling system according to the present invention, and yields the information equivalent to that obtained from the Southern blotting technique.
  • Automation of a DNA diagnostic application for sex typing using apparatus according to the invention is described, involving detection of a repeat sequence in the DYZI locus on the Y chromosome (11).
  • the repeat unit length is 3.6 kb, and anywhere from tens to thousands of the repeat units may be present in tandem depending on the nature of the DNA sample.
  • the usefulness of this Y-chromosome repeat detection for the clinical chemist lies in its ability to identify quickly the presence of male DNA in unknown samples. It can serve both as an initial screening before further expensive testing or simply as a positive control in forensic or X-linked genetic disease testing.
  • a single Eco RI repeat from this genetic region has been cloned into a plasmid vector and used successfully as a hybridization probe to detect the presence of male DNA (12). Detection of this Y-chromosome repeat is typically done using the conventional Southern blotting procedure.
  • the unlabeled oligonucleotide used for probe labelling (“Rsa I ligaid") has sequence 5' TCA ACA TCA TAA CIG AAA A 3' and is diluted to a final concentration of 5 pmol/micro-L.
  • a 60 base length oligonucleotide containing ten fluorescein molecules (“[F]60mer”) is prepared from the sequence 5' CTT TTC TTT TCT TTT CTT TTC TTT TCT TTT CTT TTC TTT TCT TTT CAG TTA TGA TGT TGT 3' and is used for target labelling.
  • the unlabeled oligonucleotide is reacted with metabisulfite/EDTA to modify citosine residues for attachment with 6-Methyl-fluorescein- N-hydroxysuccinimide ester (18).
  • the product is HPLC purified and its concentration determined spectrophotometrically by a ratio of dye to DNA absorption (19).
  • a biotin labelled oligonucleotide (“[B]30mer”) is used for probe labelling. It is synthesized in the same fashion as described above with the sequence 5" TXX XTT TTT TTT TTA GTT ATG ATG TTG T 3' where X represents modified cytosine residues which contain an amino linker arm (Molecular Biosystems, San Diego, CA). After purification and quantitation, the oligonucleotide is reacted with biotin-N-hydroxysuccinimide ester (Pierce, Rockford, IL) and purified by HPLC in a manner analogous to the fluorescent labelled oligonucleotide above.
  • Denaturation reagent is prepared just prior to use by mixing together 6 parts of reagent D a plus one part of reagent D b .
  • Reagent D a is composed of 200 mmol/L sodium hydroxide, and 800 mmol/L sodium carbonate.
  • Reagent D b is composed of 12.9% sodium polyacrylate, 5.85 mol/L sodium perchlorate, 10 mmol/L trisodium- EDTA, and is prepared by combining 18 mL (24.2g) of stock sodium polyacrylate, 39 mL (64.4g) of 9 mol/L sodium perchlorate (Aldrich Chemical Co., Milwaukee, WI; #20,842-6), and 3 mL of stock trisodium-EDTA.
  • Stock (43%) sodium polyacrylate is prepared by slow addition (Caution! - Heat evolved) of 50% NaOH (wt/wt) to 250 g. polyacrylic acid (Aldrich #19,202-3) until a 1:100 dilution of a 100 micro-L aliquot is pH 8.0.
  • Stock (200 mmol/L) trisodium-EDTA is prepared by dissolving 74.4 g. (0.22 moles) disodium EDTA (International Biotechnology Inc., New HAven, CT; #70182) in 900 mL deionized water and titrating with 50% NaOH (wt/wt, approx. 10 mL) to pH 8.0 and then diluting to a total volume of IL with deionized water.
  • Streptavidin functionalized magnetic particles [Magnetic Streptavidin 446D, Advanced Magnetics Inc. (AMI), Cambridge, MA] are pre-washed twice before use at 23 degrees C.
  • a 1.5 mL microtube containing a measured aliquot of magnetic particles is first placed directly against a BioMag SeparatorTM (AMI) containing rare-earth magnets to draw all the particles to the tube's wall.
  • AMI BioMag SeparatorTM
  • the supernatant is removed from the separated particles and replaced with 500 micro-L of buffer A.
  • the solution is vortexed vigorously to ensure complete resuspension of the particles.
  • Another cycle of separation and resuspension with buffer A is performed.
  • the suspension is separated, the supernatant is discarded and the particles resuspended in a volume of buffer A equivalent to the original aliquot.
  • Probe and target DNA are labelled by the covalent attachment of a derivatized oligonucleotide to restricted plasmid or genomic DNA respectively.
  • This simultaneous restriction/ligation technique has been previously described (20,21).
  • Probe labelling is performed manually as follows. A 100 micro-L reaction volume is prepared containing a 1 mmol/L ATP (Sigma Chemical Co., St. Louis, MO.
  • [B]30mer label and Rsa I ligaid amounts used are based on 2.5x stochiometric excess of each oligonucleotide over moles of single-strand (ss) "ends" produced by restriction plasmid probe.
  • the reaction product (0.1 micro-g/micro-L, 276 fmol ss ends/micro-L) is diluted to 160 fmol ss ends/micro-L with sterile deionized water.
  • Target labelling, denaturation and hybridization, capture, and magnetic particle washing are performed automatically by the apparatus.
  • a 50 micro-L aliquot of sample genomic DNA is first pre-restricted for 2 hr at 37 degrees C. in a total reaction volume of 65.5 micro-L by addition of 6.5 micro-L 10X restriction enzyme buffer, and 40 U Eco RI restriction enzyme. All restriction fragments produced are then labelled by incubation for 2 hr at 37 degrees C. in a total reaction volume of 100 micro-L containing 1 mmol/L ATP, IX restriction enzyme buffer, 60 U EcoRI restriction enzyme, 25 pmol [F]60mer, 25 pmol EcoRI Ligaid, and 10 U T4 DNA Ligase.
  • Labelled genomic DNA is denatured and hybridized with the Y-chromosome repeat specific probe by addition of 60 micro-L denaturation reagent and 1.6 pmol of biotin labelled probe in 10 micro-L, heated to 93 degrees C. for 15 min., cooled to 48 degrees C. for 30 min., and then cooled to 37 degrees C. Specific hybrids and excess biotin labelled probe are captured onto solid phase by addition of 40 micro-L of streptavidin- paramagnetic [articles with mixing and allowed to incubate at 37 degrees C. for 10 min. Three successive washing cycles of 1) magnetic separation, 2) removal of supernate (decantation), and 3) replenishment and incubation at 53 degrees C. for 2 min.
  • buffer B is performed, followed by one cycle with buffer C at 23 degrees C.
  • the particles are heated at 42 degrees C. for 15 min. to evaporate residual fluid, then maintained at 23 degrees C. until ready to load into an electrophoresis gel.
  • agarose Biorad, Richmond, CA; High Strength Analytical Grade #162-0126
  • 0.33 g. Ficoll 0.33 g. Ficoll (Sigma, #2637)
  • 133 mL deionized water 133 mL deionized water are placed in a tared flask and boiled until the agarose is dissolved. Enough additional deionized water is added to the flask to return it to its tared weight before boiling to compensate for evaporative loss.
  • the agarose solution is cooled to 40 degrees C.
  • a 2X stock electrophoresis loading buffer 2X LB is prepared from equal volumes of 10X Studier Buffer (300mmol/L NaOH, 10 mmol/L EDTA), 1 mg./mL Dextran Sulfate (Sigma Chemical Co., St, Louis, MO, #D-8906), and 15% Ficoll (Sigma, F2637).
  • Fluorescent internal lane size standards are also prepared by the simultaneous restriction/ligation of commonly available DNAs (i.e., lambda phage, phiX174 virus, or pBR322 plasmid) by methods identical to those described for target labelling, except that the "label” oligonucleotide is derivitized with the dye "JOE" (22) which fluoresces at a longer wavelength than fluorescein and can be discriminated spectroscopically by the fluorescent scanner.
  • a typical preparation is "[JJ]lambda+pBR(HindIII. In a total volume of 100 micro-L are combined 10 micro-g. of lambda DNA, 0.9 micro- g.
  • reaction product (3.2 fmol double-stranded fragments/micro-L) is diluted to 400 amol/micro-L for use.
  • Dried magnetic particles are resuspended in 6 micro-L of an equal volume mixture of size standard and 2X LB prior to loading into an electrophoresis gel.
  • the main mechanical mechanism is a three-axis cartesian robot. At the end of its “arm” (z-axis) is placed a fixed metallic syringe needle which performs all necessary fluid aspiration and dispensation steps.
  • System plumbing comprises two syringe pumps (250 micro-L and 2.5 micro-L) drawing from a common IL sterile deionized reservoir. Effluent from both syringes is directed through a narrow diameter tube to the end of the XYZ arm.
  • a cold storage (4 degrees C) compartment provides a stable environment for enzymes and probes for up to several days.
  • a temperature regulated rack for incubations allows 96 samples to be labelled simultaneously.
  • Another temperature regulated station houses a motor-controlled rare-earth bar magnet which can separate particles from 24 samples in a batch fashion.
  • a Macintosh II t!n computer (Apple Computer, Inc., Cupertino, CA) provides the user interface for both the robotic and separate scanner instruments.
  • the robotic instrument's operations are programmed and controlled through an iconic language where pictorial representations are used to describe chemical processes (24). This approach allows easy programming and editing and is quick to learn.
  • the syntax of the programming language is inherent in its structure.
  • the robotic instrument performs all the operations necessary to perform target labelling, solution hybridization, solid phase capture and paramagnetic particle wash steps.
  • the work surface is first manually loaded with all the necessary reagents, disposable reaction tubes and sample genomic DNA (target).
  • the instrument begins operation by first distributing an aliquot from each DNA sample tube into a corresponding tube position within the incubation rack for labelling. By addition of necessary reagents, each target sample is then simultaneously restricted and fluorescently labelled. Each sample plus an aliquot of denaturant and probe is then transferred to the magnetic separation station where a defined temperature profile is executed to perform denaturization and hybridization. Streptavidin paramagnetic particles are added to each sample to capture specific hybrids, and finally, the paramagnetic particles are washed several times with a series of buffers and prepared for loading onto the fluorescent scanner.
  • samples are manually loaded into gel wells in submarine fashion and electrophoresed at 4.5 volts/cm (325 milliamps) for 4 to 7 hours with buffer circulation.
  • a 370A Sequencer (ABI) modified to accept horizontal agarose gels is used to detect migrating fluorescent molecules (25). Real-time detection is accomplished by the use of laser excitation and fluorescent detection optics which scan across the gel's width, typically at a distance of 4.0 cm. from the sample wells (22).
  • Data analysis software allows for quantitative interpretation of electrophoresis data.
  • the data can be displayed in the form of a "gel view" which presents a record of all the fluorescent molecules which have passed through the scan region.
  • This gel view appears to be a photograph of the gel, but time, rather than position is along the direction of electrophoresis.
  • data can be displayed in a chromatographic view, which is a history of the fluorescence in a particular gel lane that passes through the scan region.
  • the chromatographic view is an analog to the kind of data presented by a densitometer.
  • the robot's physical performance was evaluated in a number of ways to verify proper function of temperature regulation systems and to validate liquid handling precision and accuracy. Temperature profiles at all relevant places on the worksurface were measured. Temperatures (4-100 degrees C) achieved were reproducible. No discernable deviation or drift in the temperature of tube contents was detectable with the thermocouple arrangement used which has a resolution of 0.1 degree C. Accuracy of pipetting (1-100 micro-L) has been measured both spectrophotometrically and gravimetrically and typically found to be within 1-5% depending upon sample viscosity with a Cv of 1.0% at 1.0 micro-L (data not shown).
  • Electrophoretic detection produces, after data analysis, both a reconstructed representation of a "gel” and a chromatogram view through an electrophoresis lane.
  • a major band at 3.6 kb represents detection of hybridization to multiple copies of the Y chromosome repeat unit.
  • the profile describes measure of relative fluorescence (Y-axis) as a function of arrival time at the detector of migrating species (X-axis). Arrival time can be related to molecular size since the electrophoresis gel material produces a separation based on size. Analogous to a Southern blot experiment, observed results thus give information of amount and molecular size of a DNA fragment with a sequence complementary to a given probe.
  • the system was able to produce consistent experimental results from 24 DNA samples in 10.5 hours of elapsed time from loading samples and reagents on the robot to receiving analyzed data from the scanner.
  • the robotic instrument was routinely loaded with reagents and samples toward the end of a work day and operated overnight (actual operating time of 6.5 h).
  • the actual hands-on-time by an operator was less than two hours and required only placement of reagent tubes in holes, gel preparation and loading, and computer interaction. This system can then provide genetic information from a DNA sample overnight as compared to typically days than is currently available with manual Southern blotting.
  • the MAcintosh II tB1 controller of the robot and scanner instruments have been interfaced to an EthertalkTM network and have allowed sending both process control code and resulting data between workers.

Abstract

A liquid-handling instrument (11) has a worksurface (22) with registration for modular stations to support containers of liquid, pipette apparatus with a pipette tip (33) coupled to a sensing circuit, a robotic translation system (31) for moving the pipette tip, and a control system with an iconic user interface for programming and editing. A gauge block (24) registered on the worksurface provides for calibration using the sensing tip, and register cavities on the worksurface provide for modular stations. There is a wash station (25) for the pipette tip on the worksurface. An automated laboratory based on the liquid-handling system has heating and cooling and a sealable incubation station (21) as well as a magnetic separation station (26 and 29). Methods are disclosed using the apparatus to convey droplets of liquid, to aspirate with minimum tip contamination, to mix liquids in containers, and to validate the worksurface. A duck-billed closure (253) is disclosed for minimizing evaporation and cross-contamination during processing, and is a part of a container disclosed for storing and transporting liquids.

Description

Automated Molecular Biology Laboratory
Cross Reference to Related Applications
The present invention is related to copending international application "ROBOTIC INTERFACE", number PCT/US90/06000, by Harry A. Guiremand, filed October 16, 1990, which is hereby incorporated by reference.
Field of the Invention
The present invention is in the field of apparatus and methods for performing chemical studies and analyses and has particular application to chemistry protocols involving genetic material from samples of DNA.
Background of the Invention
There has been rapid growth in recent years in apparatus and methodology for biochemical enterprise, particularly in the development of increasingly sophisticated systems for automating biochemical processes.
Procedures in chemistry, particularly in biochemistry, present generally more difficult problems for automation than many other kinds of processes and procedures. One reason is that there is often a very long sequence of steps in a biochemical procedure, such as gene detection and sequencing DNA. Another is that an automatic system needs to be very versatile, because different kinds of starting materials and different analytical purposes require different steps, different order of steps and the use of different kinds of chemical reagents. A third is that sample quantity is, for various reasons, quite limited, and only very small volumes, often on the order of microliters, must be used. Systems have been attempted to do procedures useful in biochemical analysis, such as transfer of liquid from one container to another by pipette, and in general such systems mimic manual procedures. Typically a mechanical arm is moved over a limited area and carries one or more pipette tips. Systems of the prior art, however, have been mostly addressed to protocols in which liquid transfer is in volumes much larger then the microliter volumes often encountered in biochemical procedures, and these systems have been less than notably successful in addressing problems created by conditions such as those described above, like pipetting very small quantities of liquid with accuracy.
Aspirating liquid into and dispensing liquid from a pipette can be done several different ways. If a liquid is dispensed into air relatively rapidly, the liquid is dispensed at a regular rate, that is, in an analog fashion. If the same liquid is dispensed relatively slowly, the dispensing rate becomes, at some point, incremental. A droplet forms on the tip, grows, and separates from the tip, then another droplet grows and separates. The size of the droplet depends on such variables as the diameters and the design of the tip and the viscosity and surface tension of the liquid being dispensed. The viscosity and surface tension depend on other variables, among them the liquid material and the temperature.
The droplet phenomenon affects aspiration of liquid into a pipette from a container of liquid as well. Liquid is aspirated with a pipette below the surface of liquid in a container, but when the tip is withdrawn, a droplet can form on the tip, and affect the accuracy of the aspiration. The effect of the droplet size on accuracy depends on the volume to be aspirated and the droplet volume.
If a volume to be aspirated or dispensed is very large compared to the droplet size that forms on the pipette tip, the droplet phenomenon has little effect on accuracy. If, however, the amount to be aspirated or dispensed is in the range of, for example, ten times the volume of a single drop, the droplet phenomenon can be serious indeed, and accuracy may be seriously impaired. In the case of biochemical procedures, the sample size and the volume of material to be aspirated and dispensed is typically very small. If a liquid to be handled is quite viscous, such as genomic DNA for example, the droplet problem assumes larger proportions.
If liquid is to be dispensed into a container, and the container already contains liquid, the pipette tip can be submerged in the liquid in the container, much in the manner that liquid is typically aspirated, then additional liquid may be dispensed in an analog fashion. A new problem in this procedure, however, is that when the pipette is withdrawn from the liquid in the container, an uncontrolled amount of the liquid can adhere to the outside of the pipette and be carried away when the pipette is moved. Again, if the volume to be aspirated is large compared to the amount that adheres to the pipette, the inaccuracy is small. If the amount to be aspirated, however is small, as is typically the case in biochemical procedures, such as DNA sequencing, the amount that adheres to the outside of the pipette may introduce significant error. Also, the further a tip is immersed in a liquid whether aspirating or dispensing, the more liquid can adhere to the tip, and the greater may be the inaccuracy.
Another problem encountered is with the speed and precision of robotics. A robot for moving a pipette to accomplish liquid transfers from container to container is in some respects a simpler problem than manipulating solid objects. For example, a robot to do pipetting requires three degrees of freedom, while some robot devices require as many as seven. In biochemical procedures, however, it is generally necessary to access a large number of different sites, and to do so very accurately. It is desirable in gene detection and DNA sequencing, for example, to process a relatively large number of samples in a single procedure. To do so requires the addition of many different reagents for each sample, and the needed reagents are not in every case the same for each sample. There have to be sites in the scanned area of the robot arm for containers to hold all of the samples and for all of the necessary liquids to perform the procedures. Moreover, there is a need for other sites, such as a wash station for the pipette or pipettes and stations for such procedures as mixing, incubating, separating, and the like.
In the case of biochemical procedures, the number of sites and the lengthy procedures require that movement from site to site be accomplished quickly to save time. Moreover, the requirement for small volumes of samples and other liquids imposes a restriction of small containers, hence small targets for the pipette. Accuracy and resolution become more important for small targets.
Systems of the prior art mimic the manual processes of pipetting very poorly. A laboratory worker using a manual pipette develops detailed technique for pipetting liquids, and often employs such technique without considerable thought. For example, a worker will typically develop technique for approaching the surface of a liquid with a pipette tip very slowly, and will move the tip slowly and with precision at the liquid surface. A worker will also typically employ technique such as touching a droplet on the pipette to the surface of a liquid to transfer the droplet to the liquid mass. These movements made almost without conscious thought by a skilled worker are difficult to duplicate with a robot, and are typically not accomplished in automatic systems of the prior art.
Yet another problem encountered in automating biochemical procedures such as gene detection and DNA sequencing is associated with the systems of programming and control. It is known to operate such systems with computers and to program sequences of action for a computer to follow to accomplish the chemical procedures, but the large variation in steps, variation in variables such as heating, cooling and mixing, and the need to process a large number of samples at a time imposes a severe requirement for a system that is flexible and operator friendly, with an operator interface that is easy to use to set up process variations.
Still another problem encountered in the design of such a system is liquid integrity. Even with rapid movement of robotic components and short and compact site design, the large number of samples and large number of steps for each sample, coupled with time required for such things as heating and cooling, dictates that operations must be done over long periods, such as several hours. Given long processing times and small samples, evaporation can be a serious problem, and can cause significant uncontrolled changes in liquid concentration, introducing error. Moreover, open containers invite problems in cross-contamination. Such contamination can be from carryover with pipette operation and also from evaporation and condensation.
Another very serious problem with apparatus of the prior art is that such apparatus typically uses throw-away pipette tips, with a new tip being used for every pipette transfer. Such a system has to provide for discarding tips after use, a waste container to recieve the discarded tips, storage for a large supply of fresh tips for use, and apparatus and control schemes for making the tip changes between liquid transfers. The apparatus and extra motions result in greater error than would result if a single tip could be used. Moreover, the need for discarding a tip and loading a new tip for each liquid transfer is time consuming, making the overall processing time more than would be necessary if a single tip could be used.
What is needed is automatic robotic apparatus for doing liquid transfers with very small quantities of liquids, and in a manner that avoids carryover and evaporation. Such an instrument needs to be modular in nature so that container stations may be interchanged, with modular stations for holding containers so that such operations as sample preparation and cleaning may be done off-line. There need to be methods for operation of such apparatus that allow a relatively large number of samples to be processed at a time, with samples and reagents placed in a close array to preserve space. The robotic actions need to be rapid to minimize overall processing time and extremely accurate to be able to access many small sites. Such a system also must incorporate robotic techniques to approximate human handling of pipette tips to accomplish adequate accuracy when operating with very small volumes of samples and reagents, and also when handling viscous liquids. The apparatus needs to provide a single pipette tip that can be reused to avoid the clumsy, time-consuming, and error- prone process of frequently discarding a tip and loading a new tip, and the problems of cross-contamination caused by single tip use must be addressed. The apparatus and associated methods of operation also must minimize evaporation and cross-contamination. Such an apparatus needs to be integrated with a control system that allows an operator to easily and quickly set up procedures with different variables, different step sequences, and different samples and reagents.
Also needed is laboratory apparatus based on such a liquid handling system to incorporate further techniques, such as temperature control and a separation station, to be able to fully automate specific chemistry protocols such as for gene detection and DNA sample purification.
Summary of the Invention
In accordance with the preferred embodiments of the present invention there is provided a liquid-handling instrument to transfer liquid between containers supported on a worksurface. The instrument has a pipette system for aspirating and dispensing liquid and a robotic translation system for moving the tip of the pipette into and out of the containers. There is a washing device for washing the pipette tip between transfers of liquid to avoid cross-contamination and a control system for programming steps for liquid transfers and for controlling the instrument. The pipette system has a sensing system to sense and communicate proximity of the tip to surfaces on the instrument to the control system. In one embodiment the sensing system has a conductive tip connected to a capacitance sensor. The sensing feature lets the robotic system move the tip with the precision needed for aspirating and dispensing very small volumes of liquid.
In another embodiment there is a gaugeblock registered to the worksurface for use in calibrating the control system relative to a precise position on the worksurface. The worksurface also has registration cavities so modular stations may be substituted on the worksurface without losing position integrity, which provides for cleaning and sample setup off-line.
The instrument has two syringe pumps connected to the common tip, and the pumps have different capacities, so course and fine aspirations and dispenses may be made with the same tip.
The robot in an embodiment is a cartesian device driven by electrical drives with two directions of travel in a horizontal plane over the worksurface and a third at right angles to the surface. The control system has an iconic, user-friendly interface for a user to program steps and enter and edit variable values. The icons are arranged in a manner that more primitive icons are nested in higher-order icons such that higher-order icons can be expanded-in-place to show more program detail without losing relationship with position in a program.
A duck-billed closure is disclosed for closing a container to minimize exposure of liquid in the container while allowing easy access by a needle-like device. A liquid-handling instrument according to the invention uses containers with the duck-billed closures to help prevent cross-contamination and evaporation. A container with a duck-billed closure is also disclosed for storing and transporting liquids.
An automated laboratory of the present invention for performing chemistry protocols is based on the liquid-handling instrument and has heating and cooling systems to control temperature of samples and reagents during processing. The laboratory has a heated and cooled incubation station with coated container cavities and a latching, sealing lid for sealing container cavities while incubating. The laboratory also has a magnetic station for separating paramagnetic particles from liquids, and the magnetic station has a magnet bar moveable vertically between rows of containers of liquid.
A method is also provided to transfer discrete droplets of liquid, and another method is provided to aspirate small volumes of liquid while minimizing tip contamination. Yet another method is provided to mix liquids effeciently with apparatus according to the preferred embodiments. Still another method is provided for validating the placement of elements on a worksurface of the present invention.
Brief Description of the Drawings
Fig. 1 is a perspective view of an automated laboratory according to a preferred embodiment of the invention.
Fig. 2A is a schematic representation of hardware components of a control system in a preferred embodiment.
Fig. 2B is a schematic representation to illustrate hardware and software structure for a control system in a preferred embodiment.
Fig. 2C is an example of a partial script list as used in the control system.
Fig. 2D is a flow diagram showing the flow of primitives for a specific script command called Dispense.
Fig. 3A is a perspective view of a robotic arm assembly for movement in the horizontal plane.
Fig. 3B is a perspective view of a robotic arm assembly also for movement in the horizontal plane, but at right angles to the movement of the arm of Fig. 3A.
Fig 3C is a perspective view, partially in section of a robotic assembly for vertical movement. Fig. 3D is a perspective view in section of the vertical movement assembly showing additional detail.
Fig. 3E is a view of a conductive pipette tip in the preferred embodiment.
Fig. 4 A is a plan view of a magnetic station. Fig. 4B is an elevation view in section of the plan view of Fig. 4A with a magnet extended.
Fig. 4C is a section view similar to Fig. 4B, but with the magnet retracted.
Fig. 4D is a section view of a tube of liquid showing a pipette tip and a helical path used for mixing liquid.
Fig. 5A is a view of a computer display showing a high-level icon representing an automated chemistry protocol.
Fig. 5B is an expansion-in-place of the icon of Fig. 5A. Fig. 5C is an expansion-in-place of one of the icons of Fig. 5B. Fig. 5D is a further expansion-in-place of an icon of Fig. 5C. Fig. 5E is yet a further expansion of an icon of Fig. 5D. Fig. 6A is a schematic representation of some steps of an example chemistry protocol for the preferred embodiment.
Fig. 6B is a representation of further steps of the example protocol of Fig. 6A.
Fig. 6C is a representation of further steps of the example protocol of Fig. 6 A.
Fig. 6D is a representation of still further steps of the example protocol of Fig. 6A.
Fig. 7A is a perspective view of an assembly of a duck-billed closure to a container.
Fig. 7B is a section through the assembly of Fig. 7A. Fig. 7C is another section through the assembly of Fig. 7A at right angles to the section of Fig. 7B.
Fig. 8 is a section view through an assembly of a duck-billed closure and a container showing a pipette tip inserted through the closure.
Fig. 9A shows one step of a method for transferring a droplet of liquid with apparatus according to the preferred embodiment.
Fig. 9B shows another step of the method of Fig. 9A.
Fig. 9C shows yet another step of the method of Fig. 9A.
Fig. 9D shows still another step of the method of Fig. 9A.
Fig. 10A shows one step of a method for aspirating liquid using apparatus according to a preferred embodiment.
Fig. 10B shows another step of the method of Fig. 10A.
Fig. IOC shows yet another step of the method of Fig. 10A.
Fig. 10D shows still another step of the method of Fig. 10A.
Fig. 11 shows a section through a wash station in a preferred embodiment.
Fig. 12 shows a section through a container at an incubation station in a preferred embodiment, with a pipette tip inserted into the container cavity.
Description of the Preferred Embodiments
General Description
Fig. 1 is a perspective view of a preferred embodiment of an automated laboratory (AL) 11 for performing chemical processes involved in molecular biology. A computer 13 with a CRT monitor 15, a keyboard 17 and a mouse device 19 is connected to the AL. The computer, CRT display, mouse, and keyboard are hardware components of a control system with an operator interface for programming the AL to perform sequences of activities, for starting and stopping processes and sequences of processes and for entering and altering process variables for specific activities. In the preferred embodiment the computer is a Macintosh II CX computer made by Apple Computer of Cupertino, CA, but other computers may also be used.
In a preferred embodiment of the invention for performing DNA sequencing the AL has a closeable, heated, clamped-lid thermal cycling station 21, an actively cooled enzyme storage station 23, a wash station 25, a reagent storage position 27 for storing and presenting frequently used reagents, a DNA sample stage 28, a wash buffer storage 30, and two magnetic particle wash stations 26 and 29 for manipulating paramagnetic particles in suspension in liquid mixtures. Also shown is a gauge block 24 for use in calibrating the robotic drives for the apparatus. The various stations are arranged on a worksurface 22. Width DI of the worksurface where all of the stations are arranged is about 50 cm and depth D2 is about 35 cm. The height is about 17 cm. In the preferred embodiment the stations on the worksurface are registered in accurately machined cavities relative to the gauge block so modular stations may be interchanged while maintaining information about the position of containers relative to the worksurface.
The magnetic particle wash stations shown are not required for the DNA sequencing protocol included in the description of the preferred embodiment, but are useful for other chemistries and illustrate the flexibility of the apparatus and to provide for ability to do chemistry protocols other than the DNA mentioned above. For example, a projected use of the apparatus of the invention is in purification of biological samples, and the magnetic particle wash stations would be used. A portion of the AL at region 46 is shown cut away to better illustrate the components in the work area.
Thermal cycling station 21 has a 96 position array of reaction cavities in 8 columns and 12 rows. The representation in Fig. 1 does not show 96 stations for reasons of detail, and the number 96 is convenient, as it is compatible with the 96 well Microliter plate known and used in the industry. There can be more or fewer reaction cavities. The reaction cavities are machined into an aluminum plate that is electrically resistance heated and also has internal water cooling passages and a thermal sensor for feedback control. Temperature is controlled in the range from 4 degrees C. to 100 degrees C. in the preferred embodiment with 1 degree C. per second rate of change. The reaction cavities are coated with Paralene (TM), a largely chemically inert coating for which materials and process are available from Solid Photography, Inc. of Melville, N.Y.
A hinged lid has a polymer undersurface such that, when the lid is closed, the reaction cavities are sealed. Each reaction cavity has a machined detail ring to contact the polymer undersurface to effect sealing (see element 285, Fig. 13). The lid is closable automatically and held closed by a latch in the preferred embodiment. Clamping by the latch is necessary to effect an adequate seal on the seal ring. Various kinds of lid drives, such as motor and pneumatic drives are useful, and various kinds of latches may be used, such as mechanical or magnetic. Sealing prevents evaporation, which helps to preserve liquid volume integrity and prevent vapor cross-contamination.
Enzyme storage station 23 has three 2 by 8 position arrays for 1.5 mL screw-top tubes, such as available from Sordstadt. The block at station 23 has cooling passages for maintaining temperature of stored enzymes at 4 degrees C. with a tolerance of 1 degree C. Although not shown in Fig. 1, a top closure is provided for station 23 with holes in the same array as the 48 tube positions, and the holes are slightly larger in diameter than the pipette tip. The top closure helps to maintain the lower temperature desirable for enzyme storage and holds the tubes in place.
Wash station 25 is for washing the pipette tip between liquid transfers to avoid carryover type cross-contamination. The wash station is connected to a waste drain and serves also as a disposal station for liquids that must be expelled from a pipette in a process protocol.
Reagent storage position 27 has positions for 1.5 mL screw-top tubes and has no active heating or cooling. The number of positions is optional. Typically 48 positions are provided.
DNA sample stage 28 has 96 positions in an 8 by 12 array for tubes containing DNA samples, also with no active heating or cooling.
Magnetic particle wash stations 26 and 29 each have a 2 by 12 array for 1.5mL microtubes, and station 26 has active heating and cooling, similar to station 21. Each magnetic station has a three- position vertically moving magnet. The magnets are for manipulating paramagnetic particles used in various protocols to capture specific material from solution.
Wash buffer storage station 30 has positions for storage containers for buffer storage. Active heating is provided with temperature sensing and control.
A cartesian transport apparatus 31 moves a pipette needle 33 of a system for aspirating liquids from containers at the various stations and dispensing liquids at the same or other stations. The pipette system includes two motor-driven syringe pumps 32 and 34 in the preferred embodiment. Pump 32 is for relatively course transfer, and pump 34 is for transfer of precise amounts of liquids. Typically pump 32 has a larger capacity than pump 34, and the capacity varies depending on the application. For example, pump 32 can vary from 250 microliter capacity for some protocols to 5 mililiters for others, and pump 34 typically has a capacity from 50 to 100 times smaller than pump 34. The two syringes have a common source of diluent. In the preferred embodiment TFE tubing is used from the syringes to the pipette probe tip, with an internal volume of l.lmL. The probe is fitted with a highly polished stainless steel tip that can convey about δ microLiter maximum droplet size.
The probe tip in the preferred embodiment is made part of a sensing system for determining when the tip approaches or touches a surface. The tip is conductive, and a wire from a capacitance sensing device is connected to a an electrical contact that contacts the probe tip. A signal is provided to the control system whenever the tip contacts a surface on the AL, and with appropriate circuitry, known in the art, proximity to a surface may also be detected without actually touching. One use of the capacitance sensing tip is to sense the surface of liquids when positioning the tip for liquid transfer operations. By sensing a liquid surface and at the same time keeping track of the height of the tip relative to the worksurface, the liquid level, hence the volume of liquid in a container can be determined. Sensing a liquid surface also provides information as to when and where to aspirate and dispense liquid while minimizing tip contamination.
Another use for the sensing tip is to examine the physical nature of the working area over which the sensing tip may pass. By passing the tip over the working area at a pre- determined height, at which height the tip will encounter no obstacle if all parts are in their proper place, one can validate the working area. If the tip encounters a surface at any place a surface should not be encountered, it is known that there is a part out of position. The control system can be programmed to provide a warning in any such circumstance.
Transport device 31 moves along slot 35 passing over the storage and activity stations. The pipette needle is movable along arm 37 of the transport device in the direction of arrow 39 and the transport is movable along slot 35 in the direction of arrow 41 to position the pipette over any container position at any station. The pipette needle is translatable vertically in the direction of arrow 43 so the transport apparatus is a cartesian XYZ mechanism capable of placing the pipette in any container on the AL work surface.
A gauge block 24 in one corner of the work area is used for calibrating the control system as to position of the pipette tip. The gauge block and the active sites on the work area are all pinned to the worksurface with accurate known dimensions. The stations on the worksurface are modular in this fashion, such that a station can be easily and quickly removed and another put in its place, or one kind of station may be substituted for another on the worksurface. Making the stations modular and providing accurate registration to the worksurface allows accurate calibration of the robotic elements to workstation positions at all times.
The gauge block has a machined surface for each of the three directions of movement of the cartesian robot, and by approaching and sensing each of the three surfaces in turn with the capacitance sensing probe tip, an accurate home position is communicated to the control system at the start of each protocol in the preferred embodiment. The probe tip can be used in the same way to validate positions of stations and elements on the worksurface. As an example, if a tube at a particular site is wedged out of position in a register opening, such as at too great a height above the worksurface, the probe with capacitance sensing can be used to determine that fact and communicate it to the control, which may then signal for appropriate action.
The pipette is for aspirating liquid from any one container and dispensing it into any another container. With the pipette, mixtures of various liquids are made and transported to any other container on the AL. The pipette system also serves to agitate liquids in a container to accomplish mixing, by repeated aspirating and dispensing of the liquid in a container, and in some instances by programmed movement of the tip in concert with aspiration and dispensing. Wash station 25 is for washing the pipette tip to avoid cross-contamination.
Computer 13, CRT 15, mouse 19 and keyboard 17 are used with the ROBOTIC INTERFACE referenced earlier, which is a unique iconic program, hereinafter called Popframes, to prepare control sequences and establish specific characteristics for the various activities that make up a complete control sequence, as well as to initiate and terminate specific strings of activities. Entries are also made at the computer to relate specific positions at specific stations on the worksurface with specific samples, such as DNA samples, and with specific reagents that are to be stored at specific sites. The iconic control program is described in further detail in another portion of this specification titled "ROBOTIC INTERFACE".
Control Functions
Fig. 2A is a block diagram showing control activities and modules in the preferred embodiment. There are many other control configurations that could be used. Computer 13, keyboard 17, mouse 19, and display 15 are connected together in the usual way, and the computer is connected by communication line 47 to a Motorola 68010 Controller PCB 51 located within the AL chassis represented by dotted enclosure 49.
Controller PCB 51 accomplishes high level control functions, such as calculations of robot position and interpretation of communication from the computer, and translation of the computer communication into more fundamental control signals for other control hardware.
The controller PCB communicates by path 53 with Function PCB 55. The function PCB accomplishes, among other functions, all of the Input/Output (I/O) operations necessary in the control operations. There are, for example, sensors on the AL to sense positions of the robot arm, such as mechanical switches. For practical reasons the sensors are operated with AC power and at a higher voltage than could be tolerated by the computer. The Function PCB monitors the status of position sensors as digital data and converts that data to computer level signals for the computer part of the control system.
In addition to the digital I\0 data described above, the Function PCB monitors analog data communicated by analog sensors on the AL, such as temperature monitoring sensors. The Function PCB converts the analog data to data suitable for the computer portion of the control system. The Function PCB handles all analog-to-digital (A/D) conversion and digital-to-analog (D/A) conversion between the computer portion and actuators and other equipment on the AL.
Function PCB 55 communicates along path 57 with the X-Y-Z robot 59, the station modules 61 on the worksurface, the syringe pumps 63 and the capacitance sensor probe 65, and also with Power Driver PCB 67 through path 69. Communications along path 57 are primarily sensor data sent to the Function PCB. Signals along path 69 are primarily signals from the Function PCB to the Power Driver PCB to actuate motions on the AL.
An AC Input and Power Supply chassis 54 in the AL receives primary AC power from outside the AL, and has the purpose of dividing, conditioning, and providing power to all the power requirements on the AL, which it does by virtue of on-board power supplies connected to the Power Driver PCB along path 56. The Power Driver PCB has the primary function in the preferred embodiment of switching power to various drivers on the PCB as required for operation, such as to the DC motors that operate the X, Y, and Z motions of the robot. The power to the various parts of the AL is provided primarily along path 58.
Fig. 2B is a largely schematic drawing to illustrate in greater detail how communication passes from the computer, a Macintosh II CX in the preferred embodiment, to other control hardware, and to illustrate in more detail the structure of software for accomplishing the tasks. As mentioned above, and described in more detail below, the interface for setup of the AL regarding constants and variable values, and for programming protocols, is an iconic program called Popframes.
In Popframes, a high level sequence of more basic steps is indicated on monitor 301 of the control computer by an icon such as icon 303. The lines 305 and 307 extending from the icon indicate sequential connection to other icons in a programmed protocol, although other such icons are not shown in Fig. 2B.
Each icon developed for Popframes is associated with a command list called a script, and the script for icon 303 is represented in Fig. 2B by enclosure 311. When an icon is activated, as in sequential performance of a series of icons to perform a protocol, the script for the icon is called in the Macintosh hardware. The script is sent to the Motorola 68010 PCB in the AL chassis in the preferred embodiment. Fig. 20 is a short excerpt from a script list. Script is programming protocol available from Apple Computer of Cupertino, CA. and used with Apple computer hardware.
The script sent to the Motorola 68010 microprocessor in the preferred embodiment is interpreted there into Forth protocols that are themselves lists of more primitive functions for the AL. For each script step there is a Forth kernel programmed on the 68010, and kernel list 313 shows a selected few of the kernels. Each script step activates a Forth kernel, and a series of primitives is performed in an order often determined by setting of flags and other variables. Communication from the Forth kernels to discrete actuators on the AL is not shown in Fig. 2B. Forth is a well known language often used in the art to program controls for robotic devices, and there are many reference books in the art explaining the structure and use of Forth.
Fig. 2D is a flow diagram showing a sequence of more primitive functions associated with one script step called Dispense, which controls dispensing of liquid from the pipette tip. Element 315, the Dispense Script command is the start of the sequence, and there are several decision points, based upon flags that can be set. One such is decision point 317, asking if the KissOff flag is set. If the flag is set, the procedure follows one path, and if not, another path is followed.
Within the sequence for Dispense the expressions enclosed in single quotes are values stored in memory that the software accesses and uses to actuate specific functions for which there may be a choice. 'descendSpeed' for example is a rate of travel for the system to use to move the pipette tip downward toward a liquid surface. As in common in the Forth language, many of the primitives are themselves combinations of even more basic functions. For example, element 319, "move down to 'dispenseLevel' at full speed is composed of a sequence that starts the vertical drive, ramps it up to full speed (pre-programmed), ramps it down near the 'dispenseLevel', and stops the drive with the pipette tip at 'dispenseLevel'.
The Cartesian Robot
Fig. 3A is a perspective view of mechanisms for driving cartesian robot 31 in the X-direction, which is the direction of arrow 41 in Fig. 1. The view of Fig. 3A has the Y-direction and Z- direction mechanisms removed, so the X-direction mechanisms may be better illustrated.
X-direction motion is provided by a D.C motor 119 that drives a flexible gear belt 121. A cast frame 123 supports the X-direction drive assembly, and the frame is mounted by conventional fasteners to baseplate 125, which is the baseplate to which stations on the worksurface in Fig. 1 are mounted. The frame is positioned precisely on the baseplate by locator pins, such as pins 127 and 129.
Motor 119 is mounted to frame 123, and a pully 131 on the motor shaft drives an intermediate toothed gear belt 133 which in turn drives another pulley 135. Pully 135 is mounted on a shaft through frame 123 in bearings (not shown) and drives yet another pulley 137. Gear belt 121 extends between driven pulley 137 and an idler pulley 139 at a distance greater than the maximum X- direction movement, which is about 45 cm. in the preferred embodiment.
A travelling cast carriage 141 is mounted below gear belt 121 on linear bearings arranged such that the carriage rides on a linear guide bar 145, which is fastened also to frame 123. Carriage 141 is attached to one side of gear belt 121 by a clamp 147 such that, as motor 119 causes belt 121 to traverse, the carriage is caused to traverse along bar 145 in the X-direction. Extension 149 from carriage 141 carries optical sensors 151 for sensing flags (not shown) fastened to the AL frame to signal position to the AL control system. The linear bearings are precision bearrings such that the maximum runout from end-to- end does not exceed about .005 inches (.013 cm).
A serious problem with previous cartesian mechanisms for liquid transfers for chemistry protocols is that the resolution and repeatability has not been sufficient for accurate probe tip placement in small vials and at closely arrayed reaction cavity positions. In the present invention, the reaction cavities, for example, at station 21 (Fig. 1) are on about 1 cm centers, and the diameter of each cavitity at the base is about .12 cm. The shaft encoders and bearings used for the X-drive, along with the control system, provide resolution of the robot in the X-direction of .020 mm.
Lands 153, 155, 157 and 159 on carriage 141 are machined at a constant height to mount mechanism for Y-direction translation. Fig. 3B shows the Y-direction mechanisms. In Fig. 3B, base plate 161 is the frame for mounting other components, and plate 161 mounts to carriage 141 of Fig. 3A and travels with that carriage. Surface 163 mounts to land 153 of Fig. 3A and surface 165 mounts to land 157 of Fig. 3A. The surfaces that mount to lands 155 and 159 on carriage 141 are not seen in Fig. 3B. Mounting plate 161 is shown as a flat plate for simplicicty, but is typically a casting with reinforcement ribs and the like in the preferred embodiment.
Y-drive motion is provided by a D.C. drive motor 167 mounted to a stand 169, that is fastened to plate 161. The motor drives a pulley (not shown) on the motor shaft, which drives a gear belt 171 around an idler pulley 173 rotatably mounted to a standoff near the end of plate 161 opposite the end where the drive motor is mounted. A moving carriage 177 is mounted on linear bearing 179 and constrained to guide along a guide bar 181 affixed to plate 161. Carriage 177 is fastened to belt 171 by a clamp (not shown) similar to clamp 147 of Fig. 3A, such that as motor 167 turns and belt 171 is driven, carriage 177 moves along guide bar 181 in the Y-direction.
Although not shown in Fig. 3B, there are optical sensors in the preferred embodiment to signal positions of the Y-direction mechanism to the control system. A Z-direction mechanism 183 is mounted to a guide bar 185 and constrained to guide in a linear bearing 187 mounted to carriage 177 to provide motion in the vertical, or Z-direction. The Z-direction mechanism is driven by a D.C. motor 189 mounted to carriage 177 and turning a pinion 191 which in turn drives a rack 193 that is fastened to the Z-direction mechanism. The Z-direction mechanism protrudes through a slot 195 in plate 161. Fig. 3C shows additional detail of the Z-direction mechanism.
Block 197 of Z-drive mechanism 183 serves as a frame for other components. Rack 193 and the guide bar for the vertical guide linear bearing mechanism are attached to block 197. A probe assembly 199 with an outer body 213 is slidably engaged in a multi- diameter cylindrical bore 201 of the body with clearance for a coil spring 203. The bore diameter is smaller at regions 205 and 207, such that the clearance between the outside of body 213 and the guide diameters of the bore is about .1 mm., while the clearance in the region for the coil spring is about 1mm. Spring 203 is captured between a shoulder 209 in block 213 and a shoulder 211 on body 213.
Body 213 is limited in vertical travel by shoulder 215 in block 197 and shoulder 211 on body 213. The vertical travel against the spring is for sensing contact with a resisting surface without damaging probe tip 33. Although not shown in Fig. 3C, there is a flag and optical sensor associated with the mechanism that signals when body 213 is lifted against spring 203.
Block 197 and body 213 in the preferred embodiment are made of an engineering plastic material to be non-conductive, such as nylon. There are several suitable materials. Tip 33 is stainless steel and brazed in the preferred embodiment to a stainless steel cylinder 217 which fits in a bore in body 213. A stainless steel thumb nut 219 threads onto body 213 and captures cylinder 217. A probe contact 221 connected to wire 223 supplies electrical potential to the probe tip, and is captured between thumb nut 219 and a thumb screw 225. Non-conductive polymer tubing 227 leads from the probe tip to the syringe pumps.
Fig. 3D is a vertical section view of the probe assembly shown without the coil spring, contact 221 and the electrical wire. Body 213 in vertical section is shown engaged in block 197 with stainless steel cylinder 217 captured in a bore in body 213 by thumb nut 219 which engages body 213 by threads 229. Thumb screw 225 is shown threaded into thumb nut 219 by threads 231. The contact, which is captured between the thumb nut and thumb screw is not shown. Ferrule 233 is a separate piece for establishing a seal between the probe tip assembly and the delivery tubing by virtue of pressure applied with the thumb nut. Although the coil spring is not shown in Fig. 3D, an optical sensor 235 is shown that senses movement of body 213 in block 197.
Further detail of the probe tip is shown in drawing 3E. Probe tip 33 is part of a brazed assembly including stainless steel cylinder 217. Overall length D7 in the preferred embodiment is about 87 mm and the length D6 of cylinder 217 is about 20 mm. The diameter D5 of cylinder 217 is about 6.4 mm (.25 inches). The tube portion is made of type 304 stainless steel tubing of about 1.27 mm (.050 inch) outside diameter and about .8 mm (.032 inch) inside diameter. For a length D3 at the tip end of about 6.4 mm (.25 inches) the tube is narrowed so the inside diameter D4 is about .3 mm (.012 inches). Having the diameter at the small dimension for only the tip end length is an advantage in that the flow resistance of the entire tube length is unaffected.
In the preferred embodiment the resolution in the X-direction is about .020mm, in the Y-direction about .025 mm, and in the Z- direction about .015 mm. The control system in the preferred embodiment also provides speed ramping that can be varied by an operator through the unique operator interface, and capability to program special motions, such as a helical motion in the Z- direction to facilitate mixing operations. Such special motions are implemented as combinations of two or more of the basic X, Y, and Z motions.
The cartesian robot has a home position in the back, left corner of the work area (facing the AL), with the vertical drive at the full up position. This home position is determined by optical sensors built into each of the three direction mechanisms. For more accuracy than is possible with the optical sensor, a home position protocol is programmed in which the tip is moved slowly to touch each of three reference surfaces on a gauge block (block 24 in Fig. 1), and the robot position is recorded for each of the three points at the time that that capacitance sensing tip touches each of the three reference surfaces. This protocol is performed typically each time a new chemistry protocol is commenced.
Magnetic Separation
In chemistry protocols of the sort for which the present invention is intended there is often a need to separate material of one sort from other materials in a liquid sample. An example is in the purification of DNA samples to be sequenced. One way to accomplish separation in many instances is by use of paramagnetic particles coated with a substance with an affinity for the product of interest of the chemistry protocol. For example, such separation can be particularly useful in the context of ligand receptor binding, such as with biotin-avidin complexes.
In the AL, to accomplish this kind of separation, solutions to be separated are transferred to vials at one of the magnetic wash stations 26 or 29 (Fig. 1). The use of one or the other depends on whether heating or cooling during separation and washing is known to facilitate the process. Precoated particles suspended in a buffer solution are aspirated from a position at one of the reagent storage stations and dispensed into the solutions to be processed at the magnetic wash station.
Fig. 4A is a plan view of wash station 26, with heating and cooling capability. There are two rows of twelve tube positions each at the station. In the space between the rows of tubes there is a magnetic bar 237. Fig. 4B shows a section through the station of Fig. 4A taken along section line 4B-4B. Magnetic bar 237 is attached by connector 239 through a screw mechanism (not shown) to a D.C. motor 241. The motor is driven by the control system to move the magnet vertically between the rows of tubes, in the direction of arrow 243.
In the preferred embodiment the magnets used are composed of rare earth materials, for example, an alloy of Niobium and Boron with iron, to obtain a high strength magnetic field. The field strength in the area of the inside of the tubes is about 35 million gauss-oersteds.
In a typical sequence for separation and washing the magnets are raised after the paramagnetic particle suspension is added, and the particles are attracted into closely packed regions that are eventually located near the bottom of the tubes as shown by regions 245 and 247. The ability to move the magnetic bar for the full height of solution in the tubes and to stop it at various points allows the entire solution volume to be swept by the intense field and the particles to be collected into a small area efficiently. It is also advantageous to use a long tube with a small diameter as opposed to a shorter tube with a larger diameter, because the paramagnetic particles have a shorter distance to travel through liquid to be collected. By slowly lowering the magnetic bar the collected particles are moved to the bottom of the tube.
After moving the particles to the bottom, typically the remaining solution is drawn off and transferred to a waste container or discarded at the wash station to waste. It is not possible, however, to aspirate all of the liquid in the tube, leaving only the particles and the adhered product. To avoid contamination three wash cycles are typically accomplished.
For a wash cycle the magnetic bar is withdrawn to a lower position where the field from the bar will not effect the particles in suspension, as shown in Fig. 4C. The cycle starts with the particles at the bottom of a tube as shown by position 251 in Fig. 4D. Then wash buffer is aspirated at station 30 and dispensed into each of the tubes at the magnetic wash station where separation is being done.
Typically, to help re-suspend the particles, the wash buffer is added with a programmed helical motion from near the bottom of a tube until all of the buffer is added, imparting a stirring action as the buffer is added. Fig. 4D shows a vertical section of one of the tubes of Fig. 4C and the pipette tip of the AL. The helical motion of the tip while dispensing wash buffer is approximated by path 249, and is pre-programmed using motions in all three directions X, Y, and Z. After adding the buffer, if another wash cycle is programmed, the magnetic bar is raised again to re-collect the particles. The action can be repeated as often as necessary, and is typically done four or five times.
Robotic Interface
A unique program is run on the computer in the preferred embodiment to create control programs, enter and edit variable values, and to initiate and terminate process sequences. The program, hereinafter called Popframes, is an iconic program that employs graphic symbols called icons to represent processes, process steps, and other activities, and is described in copending patent application entitled ROBOTIC INTERFACE, Serial No. 07/423,785 referenced earlier. Popframes provides a unique user interface that is useful for handling hierarchical information and for controlling many kinds of process machines and equipment.
Popframes has a set of routines allowing a user to select icons representing various activities and to organize the icons into flow schematics representing process flow, with the icons connected on the display with lines. The icons may also be nested such that a relatively complex sequence of activities may be represented by a single icon, and the single icon may be expanded in place to show a connected sequence of icons representing steps in the more complex sequence. The second level icons may also consist of sequences of other icons, also expandable in place, until, at the lowest level, icons represent fundamental process steps. The fundamental steps in the preferred embodiment are typically themselves sequences of even more basic activities. For example, a fundamental step may be a direction by the program to the AL to send the robot arm to a specific position at the DNA stage, station 28 in Fig. 1. The command from the computer to the electronics interface is equivalent to "Go to position X at station 28." The position is a known site to the control system, and sensors tell the control system where the robot arm is before the move. Quick calculation determines the magnitude of the X, Y and Z moves to reach the destination from the starting point. The system then accomplishes the necessary drive sequence with default acceleration and velocity.
Fig. 5A shows a screen display 69 in Popframes with a program icon 71 for the Taq DNA sequencing protocol. The single icon represents all of the steps and procedures of the protocol of sequencing DNA templates by the Taq procedure described above. A screen cursor 73 is movable over the area of the screen by moving mouse device 19 over a surface. This is a phenomenon very familiar to those skilled in computer arts.
By placing the cursor at the top-level Taq icon and pressing a button on the mouse twice, a procedure known in the art as "double clicking", a user can expand the Taq icon to see other icons representing more detail of the Taq DNA sequencing procedure.
Fig 5B shows the result of expanding the Taq icon in place. There are then eight icons shown in an orderly sequence representing eight sequential parts of the overall procedure. The eight are: Load 75, Setup 77, Anneal 79, Transfer d/dd 81, Transfer Taq 83, Incubate 85, Pool 87, and Shutdown 89. The Taq program icon is represented in the expansion by a box 91 surrounding the eight icons shown in sequence. There is a hierarchical relationship between the original icon, which is at the top of the hierarchy, and the sequence of eight icons of Fig. 5B, which are at one level below the top level icon. The labeling of the surround box: Taq, preserves the relationship so information is not lost.
A user can reverse the expansion process, collapsing a sequence of icons into a higher level icon. The method is by clicking on the close box 93 at the upper left corner of the Taq box within which the eight icons appear. Clicking means that the cursor is moved to the close box, and the mouse button is pressed once. The expansion then collapses back to the original Taq icon at the highest level. The highest level icon does not have a close box, because none is needed, but boxes at all levels below the highest level do have close boxes.
Fig. 5C shows the expansion result initiated by double clicking on the Incubate icon in Fig. 5B. After expansion, the Incubate process is seen to be composed of two distinct steps, step 95 to cycle 10 minutes at 70 degrees C, and step 97, which cycles the temperature after 10 minutes at 70 degrees C. to 10 degrees C. In Fig. 5C the Incubate icon has become the surround box 103 with a close box 101. The heirarchical relationship of the entire program is still preserved.
The expansion of step 95 by double clicking illustrates yet another feature of the iconic progam in the preferred embodiment. Fig. 5D shows the expansion of step 95 as a variable-entry box 105. Box 105 is at the lowest level of the heirarchical relationships in the iconic scheme, and provides several text fields for entering information for the computer to follow when performing the step. Rack entry field 107 allows a user to enter the name of the rack where the temperature cycling is to be done.
A user makes an entry by clicking on the text field, which enables the field for entry, then entering the designation of the rack from the keyboard. The entry field, while entry is being made, works much like a word processor. If a mistake is made, the backspace key allows the user to correct the error.
Temperature field 109 is for setting the temperature for the temperature step. Ramp field 111 is for setting a ramp rate for changing the temperature. Hold field 113 is for entering a time for holding the temperature at the set temperature. Failsafe field 115 is for entering a temperature range for deviation from the set variables without aborting the process.
At the point in expansion illustrated in Fig. 5D, the expansion has become too broad to be shown on the screen, and the Taq surround box shows terminated at the right edge of the screen. By placing the cursor inside the Taq surround box, holding down the mouse button and moving the mouse, a user can move the display to show the hidden portion at the right. This is a process called panning in the art. By panning a user can still see all of an expanded program, so information about the heirarchical relationships of the program is always preserved.
The description above for the Taq sequencing protocol shows only a few of the expansions possible for that particular program. At the lowest level of expansion of each of the other icons there is a variable-entry box. For example, at the lowest expansion level of the setup box, there is a variable-entry box with fields for the user to relate specific sites at each station to specific samples and reagents that are to be loaded for the analytical sequence.
In addition to the ability by text entry fields to vary many process parameters within a particular protocol, like the Taq sequencing protocol, there is also an ability to alter the steps and the sequence of steps, and to create entirely new and different programs. Functions for program creation and alteration are listed under menu headings in a menu bar, normally hidden from the user. With an appropriate key combination the menu can be displayed. Fig. 5E shows Fig. 5D with programming function menu bar 117 displayed.
There are, in the preferred embodiment, eight drop-down menus in the menu bar, labeled Setup, Tools, Run, Special, View, Edit, File and another headed by an Apple icon. The functions of these menus are further described in the co-pending ROBOTIC INTERFACE specification.
Procedure Example
Figures 6A, B, C and D illustrate a typical biochemical procedure performed on the AL in the preferred embodiment, and is illustrated both as an example of use and as a basis for further description of apparatus and methods in preferred embodiments of the invention. The example illustrated is a proprietary Applied Biosystems, Inc., protocol based on the Sanger termination method for DNA sequencing with Taq polymerase, performed on one single- stranded DNA template.
Each column in Fig. 6A, 6B, 6C and 6D represents one step in an automated protocol, with the progression of steps numbered at the top of the columns, reading from the left to the right through the four figures. The liquid volume dispensed to a container in any operation is listed to the right of the container, and the total liquid volume in the container is in parentheses. The protocol involves only three mechanical functions in the automated system: robotic positioning of the pipette tip, small-volume liquid handling through and with the pipette tip, and heating and cooling.
The user begins the chemistry by loading tubes of the DNA template to be sequenced and the necessary reagents in the robotic system. The DNA sample tubes are loaded to station 28 (Fig. 1), the DNA stage in the preferred embodiment. In the particular protocol illustrated there is a requirement for four samples of the same DNA template. Typically, several different DNA templates would be sequenced, and the 96 position array at the DNA stage in the preferred embodiment allows 24 different templates to be sequenced at the same time.
In the Popframes software system used to control the AL in the preferred embodiment there is facility to relate specific sites at specific stations with DNA templates and reagents, so the system "knows" where to find templates and reagents, and there is facility also, for programming sequences such as the Taq gene scanning sequence described, so the system "knows" what steps to perform in what order. It is assumed in this example that the programming has been done for Taq sequencing.
In steps 1 through 6 dye-labeled primers are annealed to the DNA template. In step 1 DNA template is moved from the DNA stage to containers at the thermal cycling station 23 (Fig. 1). One template is prepared for each of the four base types A, C, G and T. Taq sequencing buffer is moved in step 2 from reagent storage station 27 to the containers at the thermal cycling station in the amounts shown in the figure. In step 3 the dye-labeled primers are added, and in step 4 pure water is added to each reaction container.
At step 5 the lid is closed at station 21, heat is applied, and the dye-labeled primers are annealed to the DNA templates at 55 degrees C. for 5 minutes. In step 6 the reaction containers are cooled at 20 degrees C. for 20 minutes. In steps 7-14 the Taq DNA polymerase synthesizes complimentary DNA chains along the DNA templates to the dideoxynucleotide terminations. The Taq enzyme is deactivated in the alcohol precipitation of steps 15-18. At step 18 the product is ready for flourescent sequencing by gel electrophoresis.
In the process described here for 24 templates processed in parallel, the robot makes 751 moves, or 31 moves per template. For laboratories that process up to hundreds of samples per week, the number of necessary moves provides motivation for automating the protocol. In this particular protocol the magnetic wash stations are not needed, but they are useful in other protocols, such as gene scanning.
Another example of a specific molecular biology processes that the apparatus has been used to perform is provided in the section of this specification titled "Appendix A - A Further Application Example". The examples presented are not intended to limit the application of the apparatus, which is useful for many other procedures in chemistry. Applications comprise automated specific gene detection, automated nucleic acid sequence detection, and automated fluorescent labelling of nucleic acids, among other procedures.
Liquid Handling
All of the robotics in the AL are involved with handling of small volumes of liquid to accomplish chemistry protocols. Some of the liquids are quite vicous, such as genomic DNA. Others are much less viscous, such as water. A significant difference from previous equipment is in the fact that the AL of the invention uses a single pipette tip rather than throw-away pipettes as is typical in previous machines. Also in the preferred embodiment unique equipment and methods are employed to reduce evaporation to a minimum and to facilitate handling of samples and reagents to and from the AL.
Fig. 7A is a perspective view of a tube closure 253 used in the invention to prevent evaporation of materials during processing, and to provide other advantages. Closure 253 is called a duck¬ billed closure, and is shown assembled to a tube 255 of a sort often used for samples, enzymes and reagents. Closure 253 is molded from a flexible material, typically butyl rubber in the preferred embodiment. Such duck-billed closures are a feature useful in many, but not necessarily all, applications of the present invention. The closures are most useful in embodiments where problems related to evaporation are potentially more serious than in other applications.
Fig. 7B is a vertical section of the tube and closure shown in Fig. 7A along the section line 7B-7B. Fig. 7C is a vertical section of the same assembly along the section line 7C-7C, taken at a right angle to section 7B-7B.
The duck-billed closure in the preferred embodiment has a seal portion 252 with a cavity, usually circular, for enclosing the upper rim of a container to be closed. There is a flexible duck¬ billed portion 254 extending into the container from above, such that a needle-like device, such as the probe tip in the preferred embodiment, may be easily inserted from above and withdrawn to access liquid in the container. When the probe tip is inserted, the duck-billed closure remains urged against the tip with a bare minimum of opening for possible escape of liquid or vapor. When the tip is withdrawn, the duck-billed closure closes, and effectively prevents liquid or vapor escape.
The outside diameter D8 of the closure in the illustrated embodiment is about 13.2 mm. Dimension D9, the width of the duckbill portion is about 4.8mm. The height D10 of the duckbill closure is about 7.1 mm. The included angle Al of the duckbill portion is about 45 degrees. The wall thickness Dll of the duckbill portion is about ,25mm. These dimensions are for a closure for a particular tube, and will vary depending on the tube to be closed. Other embodiments will have different dimensions.
Fig. 8 shows the tube and closure of Fig. 7A, B, and C with a probe tip 33 inserted. The tip can penetrate the closure from above with little effort and be withdrawn with little effort as well. In penetration or withdrawal there is no mechanism or motion involved more than is involved if there is no closure at all, and the duckbill is caused to open only the exact amount needed to admit the probe. In other closure schemes, such as a snap-on lid, additional mechanism and robotic control must be provided to open and close the lid for access to the contents of a tube. The duckbill closure effectively prevents evaporation, eliminating inaccuracies and cross-contamination that evaporation can cause. At stations where heat is applied the duckbill closure not only prevents evaporation, but effectively seals against small buildup of pressure inside the tube.
There are other advantages to the duckbill closure. For example, reagents and other materials used in the AL can be packaged for transport with the duckbill closure in place, avoiding need to transfer the contents from one container to another during setup of the AL for a protocol. This is useful because it is very common to use a device like the AL at one site and to prepare samples and other materials at another. Moreover, most reagents are prepared by supply houses and sold to laboratories, who seldom prepare their own. The use of a duckbill closure in the original packaging can avoid potential for error and contamination. In the process of packaging with a duckbill closure, a secondary secure cap can be applied for shipment and removed at the use site without disturbing the contents.
Another advantage of the duckbill closure is that tubes can be removed from the AL after chemistry protocol and transferred directly to a centrifuge in those cases where centrifuging is desirable.
As was explained above, handling a very small volume of liquid very accurately with a pipette is a delicate and exacting procedure. It is no simple task manually, and the difficulty of duplicating the manual procedure sufficiently accurately has been an impediment to the development of useful robotics for automating laboratory procedures. The advances of the present invention, particularly in the area of robotic position resolution and repeatability and delicacy of maneuvering, combined with accurate capacitance surface sensing, have made it possible to develop programmed techniques to accomplish very accurate liquid transfers, both aspiration and dispensing. One such technique developed for the present invention is droplet conveyance, and has been called the "kiss off" technique. It is used to avoid problems associated with droplet formation and as a technique for transferring known volumes of liquid in discrete droplets from one container to another on the AL.
Fig 9A shows the pipette tip 33 in the preferred embodiment with a droplet 257 of liquid formed on the end. The tip and droplet are shown positioned over a vial 259 containing a liquid having a surface 261. A droplet is typically formed by aspirating liquid with the pipette, then driving a syringe pump to dispense just enough liquid to cause a droplet to form. The size of the droplet is determined by such factors as the diameters and material of the pipette tip, the angle, if any, on which the tip is cut, the material to be pipetted, the volume driven by the syringe pump, the temperature, and other factors.
In the droplet conveyance technique the probe tip with a droplet on the tip is lowered to a liquid surface, and the probe tip is stopped just as the droplet touches the surface. The point at which the droplet touches the liquid surface is known by the capacitance sensing ability of the robot control. The robot waits while the droplet transfers to the liquid, a process known as confluence, then the tip is raised. Fig. 9B shows the pipette of Fig. 9A lowered toward vial 259 to the point that droplet 257 just touches liquid surface 261 in the vial.
Fig. 9C shows the situation a fraction of a second after the droplet touches the liquid surface. The droplet is merging with the liquid in the vial and is still adherent to the tip by virtue of the surface tension of the liquid. Fig. 9D shows the situation after raising the tip. The liquid surface has separated from the liquid still in the pipette tip and from the pipette tip, leaving only a small miniscus 263 at the end of the tip. The droplet conveyance technique is used in the preferred embodiment to transfer discrete volumes of liquid as small as 1 micro-liter.
The kiss-off technique is a series of movements for the AL that are programmed into a reusable sequence with an icon, and can be placed in new sequences as required using the Popframes programming interface described above.
Another liquid handling technique that has been developed in the preferred embodiment is a technique of accurately aspirating liquids with the pipette tip while minimizing contamination of the tip. The technique is particularly applicable to handling viscous liquids, which are generally more troublesome in liquid handling than are less viscous liquids. Figs. 10 A, B, C, and D show the steps used in this technique.
First, probe tip 33 is positioned over the surface 265 of a liquid to be aspirated, as shown in Fig. 10A. Next the tip is lowered to touch the surface, sensed by the capacitance sensing ability associated with the probe tip, as shown in Fig. 10B. Aspiration of a programmed amount is accomplished slowly, typically at about 1 micro-liter per second, while the tip is at the surface as shown in Fig. 10B. The rate of aspiration is set to suit the viscosity of the liquid to be aspirated. If the amount to be aspirated is quite small relative to the volume in the container, then the tip position will not have to be adjusted vertically during the aspiration. If, however, the amount to be aspirated is large enough that the position of surface 265 might change enough to cause a problem, the position of the tip can be adjusted downward during the aspiration to maintain the relationship of the tip to the liquid surface. Alternatively, after the surface position is known by the capacitance sensor, the pipette tip can be lowered a fixed small amount to penetrate the liquid surface a minimal amount before aspiration begins.
After the liquid is aspirated, the probe tip is slowly withdrawn, typically at a rate of about 1.5 mm per second. As the tip is withdrawn, initially liquid still clings to the tip as shown in Fig. IOC, and this condition varies depending on the viscosity and surface tension of the liquid. The tip and the liquid separate as withdrawal continues, as shown in Fig. 10D. The probe is then moved to wherever is required to dispense the liquid that has been aspirated. The aspiration technique is a series of movements for the AL that are programmed into a reusable sequence with an icon, and can be placed in new sequences as required using the Popframes programming interface described above.
For liquids with low viscosity, such as water, it is frequently desirable to aspirate an air gap at the pipette tip after aspirating a volume of liquid, so movement of the pipette by the robot does not cause liquid to be dislodged from the pipette.
To avoid contamination, previous robotic devices have typically relied on discarding pipette tips after a single use, and in many cases on discarding vials and other containers as well. For example, in the incubation portion of the Taq DNA sequencing protocol used as an example in this specification, materials are moved to a closable-lid incubation station where the reaction vessels are machined into a coated aluminum plate. One of the reasons for having the reaction vessels machined into the plate is to provide a good heat transfer path to the liquid material to be heated in a reaction vessel. In prior devices the possibility of contamination is handled by throw-away liners or disposable reaction vessles, but disposable vessles lead to variability in heating and cooling.
The use of disposable pipette tips presents more than one difficulty. Capacitance sensing for calibration, surface sensing and other purposes is rendered difficult or impossible with disposable tips, particularly plastic tips, so accuracy cannot be attained and maintained. Further, there are many transfers to be made in a useful protocol, as described above, so as many as a thousand disposable tips would have to be stored, and the ability to dipose and replace the tips has to be programmed. Moreover, the process with disposable tips requires much more time, space, mechanism, and attendant possibility of error.
In the present invention advances in robotic equipment and technique, such as capacitance surface sensing and the Popframes programming and operating interface, make it more practical and easier to operate with a single pipette tip and to wash the tip between liquid transfers. Washing is adequate to avoid contamination, in part because of the liquid handling techniques described above, which limit exposure of the exterior of the tip to the liquids being handled.
In the preferred embodiment wash station 25 is used as needed between liquid transfers to cleanse the tip before a different reagent or sample material is transferred. The tip can be washed both inside and outside. Fig. 11 shows the pipette tip in position to wash the tip at wash station 25. The wash station includes a body 279 with a fountain 271, a well 273 and a drain 275. Body 279 is shown in section so the position and nature of other components may be seen. The fountain is a generally cylindrical bore of a depth and diameter such that wash buffer dispensed from the pipette tip will backflow and wash the outside of the tip. In the preferred embodiment the tip dimensions may vary for different protocols and purposes. In one case the tip is about .6 mm in outside diameter for a length from the end of about 6 mm. For this particular tip the depth of the fountain D12 is about 6 mm and the diameter D13 about 1.2 mm. The requirement is to provide an annulus for liquid backflow around the outside of the tip to backwash the outside of the tip beyond the length that will be inserted into a liquid on the AL.
Wash buffer dispensed from the pipette tip at the wash station to cleanse the tip backflows vertically in annulus 277 and spills over into well 273, where it drains through drain 275 to a waste container below the worksurface in the AL. The wash station serves also as a waste disposal station. For waste disposal from the pipette tip without washing the tip, the tip is positioned over well 273 and the waste is dispensed to drain 275. For waste disposal it is not needed to position the tip in the fountain.
Fig. 12 is a section through one of the reaction vessels 281 at incubation station 21 with the pipette tip shown inserted into the vessel. The reaction vessel is machined with sloping sides such as side 283, a raised lip 285, and a cylindrical chamber 287. In the preferred embodiment the material for the plate is aluminum, for the desirable heat transfer characteristics, and the surface is coated with Paralene (TM) before use so the aluminum cannot react with the materials placed in the reaction volume. The Paralene coating is not shown in Fig. 12.
The raised lip is so the lid, which has a sheet of flexible material on the undersurface, butyl in the preferred embodiment, will seal to the reaction vessel when the lid is closed. Chamber 287 is where material is actually deposited and where reaction is accomplished.
The plate at station 21 into which the reaction vessels are machined is a replacable modular unit, so plates can be assembled to the AL with reaction vessels of different sizes for different purposes. The vessel shown in Fig. 12 is for reaction volumes of about 50 micro-liters. Diameter D14 is about 1.25 mm and depth D15 is about 1.52 mm. Total depth D16 is about 8 mm, diameter D17 is about 6 mm, and angle A2 is about 40 degrees in the embodiment shown.
Material is deposited in chamber 287 for reaction, and removed from the chamber when reaction is complete. Before another reaction can be accomplished with possibly different materials entirely, the chamber has to be cleaned, which is accomplished in much the same manner as the cleaning of the pipette tip at station 25 described above. First a quantity of wash buffer is aspirated at storage station 30 (Fig. 1), then the pipette tip is moved to the reaction vessel as shown in Fig. 12. Wash buffer is dispensed into chamber 287 and backflows in the annulus between the tip and the wall of the chamber, similar to the action at the wash station. The volume above chamber 287 is large enough for a relatively large volume of buffer to be used in the process. After the washing action, the residue is pipetted to waste at station 25. It has been determined in practice that the reaction chambers can be washed up to five times and reused before the plate at the incubation station has to be replaced to avoid contamination. Appendix A - A Further Application Example
There is a great need for automation in molecular biology (1), but most workers have adapted general-purpose robotic devices to the need (2), with less than satisfactory results. This application example describes a commercially important procedure performed with apparatus according to the present invention, illustrating the utility of the invention. There are many other procedures that may be accomplished with the apparatus as described, or with minor modifications. Reference numbers are provided in parentheses throughout the example, and a reference list is provided at the end of the example to provide direction further background information.
Southern blotting, a very widely practiced technique in the molecular biology laboratory, is used to determine the length of DNA fragments homologous to a particular DNA probe (3). It has proven extremely valuable in tracking genetic diseases and identifying the presence of specific forms of genes in complex samples such as human genomic DNA (4,5). The specific chemical steps required for Southern blotting (such as blotting transfer to membranes, membrane handling, and autoradiography) are not amenable to automation, though some attempts have been made (6). A novel chemistry has been developed which produces results equivalent to those from a Southern blot experiment; the process is solution based which allows for total automation by a liquid handling robot. The details of this chemistry are described elsewhere (7,8). An essential difference between Southern blotting and this new approach is that the order of the electrophoretic size separation and hybridization are reversed. The new liquid-based methodology involves the following steps: 1) genomic DNA is simultaneously digested with a restriction enzyme and fluorescently labeled; 2) the genomic DNA is denatured and a biotin labeled probe is hybridized in solution to specific target molecules within the population of restricted genomic DNA fragments; 3) the specific hybrids are captured onto the surface of streptavidin functionalized paramagnetic particles while the remainder of the restricted genomic DNA population is not; 40 non-specific genomic DNA molecules in solution and bound to the particles are removed by stringent washing; 5) the captured hybrid/paramagnetic particle complexes are loaded directly into the well of a denaturing electrophoresis gel and the released labeled target molecules are detected when they electrophoreses past a laser scanned region a defined distance from the sample loading well; 6) collected fluorescent light is measured and the resultant data is analyzed. Other workers have described techniques where hybridization precedes electrophoresis but these techniques did not produce results where the length of the fragments analyzed could be correlated exactly to fragments in a Southern blot (8,9).
The chemical methodology described above lends itself to automation with a robotic liquid handling system according to the present invention, and yields the information equivalent to that obtained from the Southern blotting technique. Automation of a DNA diagnostic application for sex typing using apparatus according to the invention is described, involving detection of a repeat sequence in the DYZI locus on the Y chromosome (11). The repeat unit length is 3.6 kb, and anywhere from tens to thousands of the repeat units may be present in tandem depending on the nature of the DNA sample. The usefulness of this Y-chromosome repeat detection for the clinical chemist lies in its ability to identify quickly the presence of male DNA in unknown samples. It can serve both as an initial screening before further expensive testing or simply as a positive control in forensic or X-linked genetic disease testing. A single Eco RI repeat from this genetic region has been cloned into a plasmid vector and used successfully as a hybridization probe to detect the presence of male DNA (12). Detection of this Y-chromosome repeat is typically done using the conventional Southern blotting procedure.
Reagents used in the Procedure
Human genomic DNA is extracted from either lymphocyte blood fraction (13) or two different harvested cell line cultures (Raji, black male; R562, Caucasian female; American Type Culture Collection, Rockville, MD (14) using a model 340A Nucleic Acid Extractor (Applied Biosystems, Inc. (ABI), Foster City, CA). Extracted DNA is dissolved in 1 mL sterile deionized water and its concentration determined spectrophotometrically (1.0 A at 260 nm = 50 micro-g/mL DNA). The DNA is diluted with sterile deionized water to a final concentration of 0.2 micro- g/micrc— L.
Oligonucleotides are synthesized by the phosphoramidite approach (15) using a Model 381A DNA Synthesizer (ABI) at 0.2 micro-mol scale (16) with (2-0-cyanoethyl)-phosphoramidites (ABI). Crude ammonia hydrozylates are purified by Oligonucleotide Purification Cartridges™ (ABI) (17), evaporated to dryness, and stock solutions are prepared by dissolving in 1 mL sterile deionized water. Oligonucleotide concentration is determined spectrophotometrically from a dilution of the stock (1.0 A at 260 nm = 33 micro-g/mL DNA).
The unlabeled oligonucleotide used for probe labelling ("Rsa I ligaid") has sequence 5' TCA ACA TCA TAA CIG AAA A 3' and is diluted to a final concentration of 5 pmol/micro-L.
A 60 base length oligonucleotide containing ten fluorescein molecules ("[F]60mer") is prepared from the sequence 5' CTT TTC TTT TCT TTT CTT TTC TTT TCT TTT CTT TTC TTT TCT TTT CAG TTA TGA TGT TGT 3' and is used for target labelling. The unlabeled oligonucleotide is reacted with metabisulfite/EDTA to modify citosine residues for attachment with 6-Methyl-fluorescein- N-hydroxysuccinimide ester (18). The product is HPLC purified and its concentration determined spectrophotometrically by a ratio of dye to DNA absorption (19).
A biotin labelled oligonucleotide ("[B]30mer") is used for probe labelling. It is synthesized in the same fashion as described above with the sequence 5" TXX XTT TTT TTT TTT TTA GTT ATG ATG TTG T 3' where X represents modified cytosine residues which contain an amino linker arm (Molecular Biosystems, San Diego, CA). After purification and quantitation, the oligonucleotide is reacted with biotin-N-hydroxysuccinimide ester (Pierce, Rockford, IL) and purified by HPLC in a manner analogous to the fluorescent labelled oligonucleotide above.
Denaturation reagent is prepared just prior to use by mixing together 6 parts of reagent Da plus one part of reagent Db. Reagent Da is composed of 200 mmol/L sodium hydroxide, and 800 mmol/L sodium carbonate. Reagent Db is composed of 12.9% sodium polyacrylate, 5.85 mol/L sodium perchlorate, 10 mmol/L trisodium- EDTA, and is prepared by combining 18 mL (24.2g) of stock sodium polyacrylate, 39 mL (64.4g) of 9 mol/L sodium perchlorate (Aldrich Chemical Co., Milwaukee, WI; #20,842-6), and 3 mL of stock trisodium-EDTA. Stock (43%) sodium polyacrylate is prepared by slow addition (Caution! - Heat evolved) of 50% NaOH (wt/wt) to 250 g. polyacrylic acid (Aldrich #19,202-3) until a 1:100 dilution of a 100 micro-L aliquot is pH 8.0. Stock (200 mmol/L) trisodium-EDTA is prepared by dissolving 74.4 g. (0.22 moles) disodium EDTA (International Biotechnology Inc., New HAven, CT; #70182) in 900 mL deionized water and titrating with 50% NaOH (wt/wt, approx. 10 mL) to pH 8.0 and then diluting to a total volume of IL with deionized water.
Three buffer solutions are used to wash paramagnetic particles and their composition is as follows: buffer A = 1.0X SSPE (180 mmol/L sodium chloride, 10 mmol/L monobasic sodium phosphate pH 7.4, 1 mmol/L EDTA), 0.5% Tween-20 (Aldrich; #27,434-8); buffer B = 118 mmol/L sodium chloride, 16.5 mmol/L sodium carbonate, 7.8 mmol/L sodium bicarbonate, 0.5% Tween-20; buffer C = lOOmmol/L sodium chloride. Streptavidin functionalized magnetic particles [Magnetic Streptavidin 446D, Advanced Magnetics Inc. (AMI), Cambridge, MA] are pre-washed twice before use at 23 degrees C. using buffer A. A 1.5 mL microtube containing a measured aliquot of magnetic particles is first placed directly against a BioMag Separator™ (AMI) containing rare-earth magnets to draw all the particles to the tube's wall. The supernatant is removed from the separated particles and replaced with 500 micro-L of buffer A. The solution is vortexed vigorously to ensure complete resuspension of the particles. Another cycle of separation and resuspension with buffer A is performed. Finally the suspension is separated, the supernatant is discarded and the particles resuspended in a volume of buffer A equivalent to the original aliquot.
Procedures
Both probe and target DNA are labelled by the covalent attachment of a derivatized oligonucleotide to restricted plasmid or genomic DNA respectively. This simultaneous restriction/ligation technique has been previously described (20,21). Probe labelling is performed manually as follows. A 100 micro-L reaction volume is prepared containing a 1 mmol/L ATP (Sigma Chemical Co., St. Louis, MO. #A-0770), 15 mmol/L dithiothreitol (Sigma, D-9779), IX restriction enzyme buffer (Promega Corp., Madison, WI), 50 micro- g/mL BSA-OAc (Promega), 10 micro-g pY3,4 plasmid DNA, 60U Rsa I restriction enzyme (all enzymes used are from Promega), 10 U T4 DNA ligase, and 69 pmol each of the [B]30mer labelled oligonucleotide and the Rsa I ligaid. Enzyme amounts used are based on units of enzyme activity per weight of DNA, using 6 U/micro-g. [B]30mer label and Rsa I ligaid amounts used are based on 2.5x stochiometric excess of each oligonucleotide over moles of single-strand (ss) "ends" produced by restriction plasmid probe. The reaction product (0.1 micro-g/micro-L, 276 fmol ss ends/micro-L) is diluted to 160 fmol ss ends/micro-L with sterile deionized water.
Target labelling, denaturation and hybridization, capture, and magnetic particle washing are performed automatically by the apparatus. For target labelling, a 50 micro-L aliquot of sample genomic DNA is first pre-restricted for 2 hr at 37 degrees C. in a total reaction volume of 65.5 micro-L by addition of 6.5 micro-L 10X restriction enzyme buffer, and 40 U Eco RI restriction enzyme. All restriction fragments produced are then labelled by incubation for 2 hr at 37 degrees C. in a total reaction volume of 100 micro-L containing 1 mmol/L ATP, IX restriction enzyme buffer, 60 U EcoRI restriction enzyme, 25 pmol [F]60mer, 25 pmol EcoRI Ligaid, and 10 U T4 DNA Ligase. Labelled genomic DNA is denatured and hybridized with the Y-chromosome repeat specific probe by addition of 60 micro-L denaturation reagent and 1.6 pmol of biotin labelled probe in 10 micro-L, heated to 93 degrees C. for 15 min., cooled to 48 degrees C. for 30 min., and then cooled to 37 degrees C. Specific hybrids and excess biotin labelled probe are captured onto solid phase by addition of 40 micro-L of streptavidin- paramagnetic [articles with mixing and allowed to incubate at 37 degrees C. for 10 min. Three successive washing cycles of 1) magnetic separation, 2) removal of supernate (decantation), and 3) replenishment and incubation at 53 degrees C. for 2 min. with buffer B are performed, followed by one cycle with buffer C at 23 degrees C. After a final magnetic separation and decantation, the particles are heated at 42 degrees C. for 15 min. to evaporate residual fluid, then maintained at 23 degrees C. until ready to load into an electrophoresis gel.
To prepare a denaturing agarose gel, 1.07 g. agarose (Biorad, Richmond, CA; High Strength Analytical Grade #162-0126), 0.33 g. Ficoll (Sigma, #2637) and 133 mL deionized water are placed in a tared flask and boiled until the agarose is dissolved. Enough additional deionized water is added to the flask to return it to its tared weight before boiling to compensate for evaporative loss. The agarose solution is cooled to 40 degrees C. by placing intermittently on ice with constant swirling, 1.33 mL of 100X Studier Buffer (3 mol/L NaOH, 100 mmol/L EDTA) is added, the solution is then cooled to 30 degrees C. and poured into aa glass bottom gel tray (22x28 cm.) and a well-forming combing introduced into the gel. 1.2 L of IX Studier Buffer is pored to cover the gel once solidified.
A 2X stock electrophoresis loading buffer 2X LB) is prepared from equal volumes of 10X Studier Buffer (300mmol/L NaOH, 10 mmol/L EDTA), 1 mg./mL Dextran Sulfate (Sigma Chemical Co., St, Louis, MO, #D-8906), and 15% Ficoll (Sigma, F2637).
Fluorescent internal lane size standards are also prepared by the simultaneous restriction/ligation of commonly available DNAs (i.e., lambda phage, phiX174 virus, or pBR322 plasmid) by methods identical to those described for target labelling, except that the "label" oligonucleotide is derivitized with the dye "JOE" (22) which fluoresces at a longer wavelength than fluorescein and can be discriminated spectroscopically by the fluorescent scanner. A typical preparation is "[JJ]lambda+pBR(HindIII. In a total volume of 100 micro-L are combined 10 micro-g. of lambda DNA, 0.9 micro- g. of pBR322-plasmid DNA, 25 pmol [j]60mer (a molecule analogous to [F]60mer but labelled with JOE) and all other reagents as indicated above for target labelling. The reaction product (3.2 fmol double-stranded fragments/micro-L) is diluted to 400 amol/micro-L for use.
Dried magnetic particles are resuspended in 6 micro-L of an equal volume mixture of size standard and 2X LB prior to loading into an electrophoresis gel.
Apparatus
The main mechanical mechanism is a three-axis cartesian robot. At the end of its "arm" (z-axis) is placed a fixed metallic syringe needle which performs all necessary fluid aspiration and dispensation steps. System plumbing comprises two syringe pumps (250 micro-L and 2.5 micro-L) drawing from a common IL sterile deionized reservoir. Effluent from both syringes is directed through a narrow diameter tube to the end of the XYZ arm.
On the work surface, a cold storage (4 degrees C) compartment provides a stable environment for enzymes and probes for up to several days. A temperature regulated rack for incubations allows 96 samples to be labelled simultaneously. Another temperature regulated station houses a motor-controlled rare-earth bar magnet which can separate particles from 24 samples in a batch fashion. There are also defined positions for twelve 1.5 mL microfuge tubes of reagent, two 35 mL bottles of buffer, five 100 mL bottles of temperature regulated buffer, a rack of 1.2 mL microfuge tube of sample DNA, and a needle tip wash station.
A Macintosh IIt!n computer (Apple Computer, Inc., Cupertino, CA) provides the user interface for both the robotic and separate scanner instruments. The robotic instrument's operations are programmed and controlled through an iconic language where pictorial representations are used to describe chemical processes (24). This approach allows easy programming and editing and is quick to learn. The syntax of the programming language is inherent in its structure.
The robotic instrument performs all the operations necessary to perform target labelling, solution hybridization, solid phase capture and paramagnetic particle wash steps. Before automatic operation begins, the work surface is first manually loaded with all the necessary reagents, disposable reaction tubes and sample genomic DNA (target). The instrument begins operation by first distributing an aliquot from each DNA sample tube into a corresponding tube position within the incubation rack for labelling. By addition of necessary reagents, each target sample is then simultaneously restricted and fluorescently labelled. Each sample plus an aliquot of denaturant and probe is then transferred to the magnetic separation station where a defined temperature profile is executed to perform denaturization and hybridization. Streptavidin paramagnetic particles are added to each sample to capture specific hybrids, and finally, the paramagnetic particles are washed several times with a series of buffers and prepared for loading onto the fluorescent scanner.
For detection of the chemical product produced by the robotic instrument, samples are manually loaded into gel wells in submarine fashion and electrophoresed at 4.5 volts/cm (325 milliamps) for 4 to 7 hours with buffer circulation. A 370A Sequencer (ABI) modified to accept horizontal agarose gels is used to detect migrating fluorescent molecules (25). Real-time detection is accomplished by the use of laser excitation and fluorescent detection optics which scan across the gel's width, typically at a distance of 4.0 cm. from the sample wells (22). Data analysis software allows for quantitative interpretation of electrophoresis data. The data can be displayed in the form of a "gel view" which presents a record of all the fluorescent molecules which have passed through the scan region. This gel view appears to be a photograph of the gel, but time, rather than position is along the direction of electrophoresis. Alternatively, data can be displayed in a chromatographic view, which is a history of the fluorescence in a particular gel lane that passes through the scan region. The chromatographic view is an analog to the kind of data presented by a densitometer.
Results and Discussion
The robot's physical performance was evaluated in a number of ways to verify proper function of temperature regulation systems and to validate liquid handling precision and accuracy. Temperature profiles at all relevant places on the worksurface were measured. Temperatures (4-100 degrees C) achieved were reproducible. No discernable deviation or drift in the temperature of tube contents was detectable with the thermocouple arrangement used which has a resolution of 0.1 degree C. Accuracy of pipetting (1-100 micro-L) has been measured both spectrophotometrically and gravimetrically and typically found to be within 1-5% depending upon sample viscosity with a Cv of 1.0% at 1.0 micro-L (data not shown).
The use of a single pipetting tip required a study of cross- contamination. Contamination resulting from carryover from one reagent tube to the next when a multiple aspiration is performed was measured spectrophotometrically to be 0.5 micro-L. Sample- to-sample cross contamination after probe tip washing is undetectable as previously reported by other workers in a similar liquid handling instrument (26). Reagent pollution due to multiple aspiration without tip washing occurs at two places in the entire process as currently practiced. The first opportunity for potential cross-contamination is in the mixing of common restriction and labelling reagents ("paletting") where the potential exists for ATP to contaminate buffer, buffer to contaminate restriction enzyme, and so forth. This poses little threat to reagent integrity since the order in which reagents are aspirated can be judiciously chosen so as to accommodate a slight amount of carry-over. Furthermore, this operation occurs only once during execution of the entire process. The second occurrence of carry¬ over due to multiple aspiration is when sample DNA is transferred from the labelling station to the magnetic separation station. Here a small amount of denaturant may contaminate the probe reagent. Probe reagent could be multi-dispensed to each tube position within the magnetic separation station before transfer of labelled DNA to alleviate this problem.
Electrophoretic detection produces, after data analysis, both a reconstructed representation of a "gel" and a chromatogram view through an electrophoresis lane. A major band at 3.6 kb represents detection of hybridization to multiple copies of the Y chromosome repeat unit. The profile describes measure of relative fluorescence (Y-axis) as a function of arrival time at the detector of migrating species (X-axis). Arrival time can be related to molecular size since the electrophoresis gel material produces a separation based on size. Analogous to a Southern blot experiment, observed results thus give information of amount and molecular size of a DNA fragment with a sequence complementary to a given probe.
Good signal uniformity (Cv=12%) was observed from the result of pY3.4 probe hybridization with five identical DNA samples.
A control experiment was done where five different DNA samples were hybridized with either the Y chromosome repeat probe or a probe which is not homologous to the human genome (plasmid pSP64 labelled in the same manner as described above). Varying signal intensity was observed within the group of male DNA's. As previously reported by Lua (12) we observed that the DNA sample of Black origin exhibited a stronger signal, and the DNA sample of Asian origin exhibited a weaker signal than the DNA sample of Caucasian origin. Also as noted in the literature, secondary hybridization to smaller size fragments was observed even in the female DNA. No hybridization with the non- homologous probe was detected in any of the human DNA samples.
The system was able to produce consistent experimental results from 24 DNA samples in 10.5 hours of elapsed time from loading samples and reagents on the robot to receiving analyzed data from the scanner. The robotic instrument was routinely loaded with reagents and samples toward the end of a work day and operated overnight (actual operating time of 6.5 h). The next morning particle suspensions e\were loaded onto the scanner instrument for electrophoresis and detection and resultant data was analyzed (4h). The actual hands-on-time by an operator was less than two hours and required only placement of reagent tubes in holes, gel preparation and loading, and computer interaction. This system can then provide genetic information from a DNA sample overnight as compared to typically days than is currently available with manual Southern blotting. Example Summary and Conclusions
From the example above it is concluded that a robotic liquid handling instrument according to the invention can be used successfully to automate specific human gene detection in such a way to yield the equivalent experimental result to that produced by Southern blotting. The manner in which this result is accomplished is simpler and faster than the manual methods typically employed. The individual liquid handling steps are executed with precision. Since operation is computer controlled the process can be performed consistently, reliably, and relentlessly providing a new opportunity for high sample throughput.
The MAcintosh IItB1 controller of the robot and scanner instruments have been interfaced to an Ethertalk™ network and have allowed sending both process control code and resulting data between workers.
The continued development of automated DNA sequencing using a robot similar to the one described herein has recently been discussed in another document (25). The robot's unique combination of attributes (accurate pipetting, XYZ motion, temperature control and magnetic particle handling) make it ideally suited to perform this and many other chemical methodologies.
References for Appendix A
(1) Landergren U, Kaiser R, Caskey C, Hood L. DNA diagnostics - molecular techniques and automation. Science 1988; 242:229-37.
(2) Wilson RK, Yuen AS, Clark SM, Spence C, Arakelain P, Jood LE. Automation of dideoxynucleotide DNA sequencing reactions using a robotic workstation. Biotechniques 1988; 6:776-87. (3) Southern, EM. Detection of specific sequences among DNA fragments separated by gel. J Mol Biol 1975; 98,3:503-17
(4) Caskey TC. Disease diagnosis by recombinant DNA methods. Science 1987; 236:1223-28.
(5) Watkins PC. Restriction fragment length polymorphism (RFLP): applications in human chromosome mapping and genetic disease research. Biotechniques 1988; 6,4:310-19.
(6) Gersten DM, Zapolski EJ, Golab TJ, Buas M, Ledley RS. Computer controlled DNA electrophoresis and hybridization. Proc. Meet Int.Electrophor. Soc. 1986; 5:187-90.
(7) Kieth D, Hoff LB, Mayrand PE, McBride LJ, Robertson J, Recknor M, Ziegel J, Meister S, Whitley N, Kronick M. Detection and sizing of fluorescently labelled DNA fragments following in- solution hybridization: an alternative to traditional Southern blotting. Manuscript submitted to Nuc. Acids Res.
(8) Kronick MN, Kieth DH, McBride LJ, Whitley NM, Hunkapiller MW. Method and kit for detecting a nucleic acid sequence. Eur. patent appl. no. 0322311, 1988.
(9) Ga per HB, Cimino GB, Isaacs ST, Ferguson M, Hearst J. Nuc. Reverse Southern hybridization. Nuc. Acids Res. 1986; 14:9943- 9954.
(10) Jones FS, Griraberg JI, Fischer SG, Ford JP. Detection of sickle-cell mutation by electrophoresis of partial RNA:DNA. Gene 1985; 39,1:77-83
(11) Bostock CJ, Gosden JR, Mitchell AR. Localisation of male- specific DNA fragment to subregion of the human Y chromosome. Nature 1978; 272:324-328.
(12) Lau Y, Schonberg S. A male-specific DNA probe detects heterochromatin sequences in a familial Yq" chromosome. Am. J. Hum. Genet. 1984; 36:1394-96.
(13) Lymphocyte preparation II. ABI 340A Nucleic Acid Extractor User Manual 1989; 3:12.
(14) Cell culture preparation. ABI Nucleic Acid Extractor User Manual 1989; 3:10.
(15) Caruthers MH, Barone AD, Beaucage SL, et al. Chemical synthesis of deoxynucleotides by the phosphoramidite approach. Methods Enzymol 1987: 154:287-313.
(16) ABI 370A User Bulletin 1986; 3:7.
(17) McBride LJ, MsCollum C, Davidson S, Efcavitch JW, Andrus A, Lombardi SJ. A new, reliable cartridge for the rapid purification of synthetic DNA. Biotechniques 1988; 6:362-7.
(18) Draper D and LE Gold. A method for linking fluorescent labels to polynucleotides: application to studies of ribosome- ribonucleic acid interactions. Biochemistry 1980; 19,9:1774-81.
(19) ABI 370A User Bulletin 1989; 11.
(20) Kieth DH, Kronick MN, McBride LJ, Whitley NM. Labelling by simultaneous ligation and restriction. Eur. patent appl. no. 0327- 429, 1989.
(21) Carrano AV, Lamerdin J, Ashworth LK. A high-resolution, fluorescence- based, semiautomated method for DNA fingerprinting. Genomics 1989; 4:129-136.
(22) Fung S, Woo SL, Menchen SM, Connel CR, Heiner C. Method of detecting electrophoretically separated oligonucleotides. Eur. patent appl. no. 0233053, 1986.
(23) Shigeura J. Mechanical Design of Small- Volume Fluid- Handling Robots for the Molecular Biology Laboratory. Proc. 5th International Symposium on Laboratory Robotics 1989.
(24) Guiremand H. Popframes programming interface. Eur. patent appl. no. 00000-00000.
(25) Connell C, Fung S, Heiner C. Automated DNA sequence analysis. Biotechniques 1987; 5:342-348.
(26) Severns ML, Brennan JE, Kline LM, Eply KM. Pipette cleaning in automated systems. J. Automatic Chemistry 1986; 8,3:135-141.
(27 ) Chem. Eng. News 1989; Nov 13:6.
It will be apparent to a worker skilled in the art that there are many changes that can be made in the details of the invention as described without departing in any significant degree from the spirit and scope of the invention. For example, the numbers of positions at the various stations need not be as shown in the preferred embodiments. More or fewer positions could be used. As another example, the dimensions and construction details can vary widely. There are many ways to accomplish the resolution needed for the robot, and many different kinds of sensors and drives that can be used. As still another example, there are many different materials that would be suitable for different parts of the apparatus, such as the plate at the incubation station, and the material described is the preferred mode. Many such changes in detail can be made without departing from the spirit and scope of the invention.

Claims

What is claimed is:
1. A liquid-handling instrument for transferring liquid from one container to another comprising: a worksurface for supporting said containers of liquid; pipette means for aspirating and dispensing liquid, said pipette means comprising a pipette tip; robotic translation means for moving said pipette tip into and out of said containers; washing means for washing said pipette tip between liquid transfers; and control means for programming steps of said liquid transfers and for controlling said instrument to perform said steps; said pipette means comprising position sensing means for sensing pipette tip position relative to proximate surfaces and for communicating said pipette tip position to said control means.
2. An instrument as in claim 1 wherein said position sensing means comprises an electrically conductive pipette tip coupled to capacitance sensing means.
3. An instrument as in claim 1 wherein said worksurface comprises a gauge block for calibrating said control means for position of said pipette tip relative to said worksurface, said gauge block being securely registered to said worksurface.
4. An instrument as in claim 3 wherein said worksurface comprises registration means for positioning a modular container station, said registration means positioned accurately relative to said gauge block, such that a first modular station may be removed from said cavity and a second modular station substituted therefor while maintaining known dimensions from containers in said stations to the known position of said gauge block.
5. An instrument as in claim 1 wherein said pipette means comprises a first syringe pump for aspirating and dispensing liquids with a first degree of accuracy and a second syringe pump for aspirating and delivering liquids with a second degree of accuracy, said first and second syringe pumps being commonly connected to said pipette tip.
6. An instrument as in claim 1 wherein said robotic translation means comprises a robot having a carriage over said worksurface for carrying said pipette tip, said carriage being translatable over the area of said worksurface and also translatable toward and away from said worksurface.
7. An instrument as in claim 6 wherein said robot is a cartesian robot having three directions of travel, two in a horizontal plane and the third direction vertical.
8. An instrument as in claim 7 wherein said robot has three principal drives, one for each of said three directions of travel, and said drives are powered by electric motors.
9. An instrument as in claim 1 wherein said control means comprises an iconic interface having user-selectable icons for programing a protocol and for entering values for control variables, said icons representing specific steps and action sequences in said protocol, said icons being expandable-in-place to show other steps that an icon comprises.
10. An instrument as in claim 1 wherein said washing means comprises a container connected to a waste disposal means, said container having a cavity closed at the lower end, said cavity being in depth at least ten times the diameter of said pipette tip, and in diameter no more than twice the diameter of said pipette tip, such that a liquid dispensed from said pipette tip with said pipette tip in said cavity will backflow in the annulus around said pipette tip for a length of said pipette tip of at least ten times the diameter of said pipette tip.
11. An instrument as in claim 1 wherein at least one of said containers supported by said worksurface comprises closure means having a flexible duck-billed portion extending toward the interior of said container.
12. An instrument as in claim 11 wherein said container comprises an upper rim and said closure means comprises an annular cavity portion surrounding said duck-billed portion for sealing to said upper rim.
13. A container for storing and transporting liquid comprising: liquid receiving means having an opening for receiving liquid; and closure means for providing closure to said opening of said liquid receiver means; said closure means comprising a flexible duck-billed portion extending toward the interior of said liquid receiver means for permitting easy entry of a needle-like device while minimizing exposure of liquid and vapor inside said liquid receiver means to external atmosphere.
14. A container as in claim 13 wherein said liquid receiving means comprises an upper rim surrounding said opening and said closure means comprises an annular cavity portion for mating to said upper rim.
15. An automated laboratory for transferring liquid chemical mixtures and solutions from one container to another and performing a chemistry protocol comprising: a worksurface for supporting said containers of liquid; pipette means for aspirating and dispensing liquid, said pipette means comprising a pipette tip; robotic translation means for moving said pipette tip into and out of said containers; washing means for washing said pipette tip between liquid transfers; heating means for heating liquids in said containers; cooling means for cooling liquids in said containers; and control means for programming steps of said chemistry protocol and for controlling said automated laboratory to perform said protocol; said pipette means comprising position sensing means for sensing pipette tip position relative to proximate surfaces and for communicating said pipette tip position to said control means.
16. An automated laboratory as in claim 15 wherein said position sensing means comprises an electrically conductive pipette tip coupled to capacitance sensing means.
17. An automated laboratory as in claim 15 further comprising an incubation station heatable by said heating means and coolable by said cooling means, said incubation station having cavities coated with a chemically inert coating, and a latching lid with a sealing surface such that with said lid closed and latched, said cavities are individually sealed.
18. An automated laboratory as in claim 17 wherein said cavities are machined into a metal block, and said metal block is registered to said worksurface and removable, such that a block of cavities may be removed and replaced.
19. An automated laboratory as in claim 17 wherein said cavities have a lower cylindrical portion for holding a sample and an upper conical portion to provide additional volume to hold liquid aspirated during a cleaning procedure.
20. An automated laboratory as in claim 17 wherein said cavities each have a machined upper lip such that force exerted on said lid by latching said lid is concentrated on said machined upper lips of said cavities to facilitate sealing.
21. An automated laboratory as in claim 15 further comprising magnetic means for passing a magnetic field through at least one container supported by said worksurface to separate paramagnetic particles from a liquid in said container.
22. An automated laboratory as in claim 21 wherein said magnetic means comprises a station supported by said worksurface, said station having two rows of containers for holding liquid, and a magnetic bar supported on an elevator such that said bar may be selectively elevated and withdrawn from between said rows of containers.
23. An automated laboratory as in claim 21 wherein said magnetic bar comprises rare-earth magnetic material.
24. An automated laboratory as in claim 15 wherein said worksurface comprises a gauge block for calibrating said control means for position of said pipette tip relative to said worksurface, said gauge block being securely registered to said worksurface.
25. An automated laboratory as in claim 24 wherein said worksurface comprises registration means for positioning a modular container station, said registration means positioned accurately relative to said gauge block, such that a first modular station may be removed from said cavity and a second modular station substituted therefor while maintaining known dimensions from containers in said stations to the known position of said gauge block.
26. An automated laboratory as in claim 15 wherein said pipette means comprises a first syringe pump for aspirating and dispensing liquids with a first degree of accuracy and a second syringe pump for aspirating and dispensing liquids with a second degree of accuracy, said first and second syringe pumps being commonly connected to said pipette tip.
27. An automated laboratory as in claim 15 wherein said robotic translation means comprises a robot having a carriage over said worksurface for carrying said pipette tip, said carriage being translatable over the area of said worksurface and also translatable toward and away from said worksurface.
28. An automated laboratory as in claim 27 wherein said robot is a cartesian robot having three directions of travel, two in a horizontal plane and the third direction vertical.
29. An automated laboratory as in claim 28 wherein said robot has three principal drives, one for each of said three directions of travel, and said drives are powered by electric motors.
30. An automated laboratory as in claim 15 wherein said control means comprises an iconic interface having user-selectable icons for programing a protocol and for entering values for control variables, said icons representing specific steps and action sequences in said protocol, said icons being expandable-in-place to show other steps that comprise an icon.
31. An automated laboratory as in claim 15 wherein said washing means comprises; a fountain for enclosing said pipette tip during washing; and a well surrounding said fountain such that liquid flowing from said fountain flows into said well, said well having a drain disposed to communicate unwanted material to an external location; said fountain comprising a cylindrical enclosure closed at the lower end, said enclosure being in depth at least ten times the diameter of said pipette tip, and in diameter no more than twice the diameter of said pipette tip, such that a liquid dispensed from said pipette tip with said pipette tip inserted in said cavity will backflow in the annulus around said pipette tip for a length of at least ten times the diameter of said pipette tip.
32. An automated laboratory as in claim 15 wherein at least one of said containers supported by said worksurface comprises closure means having a flexible duck-billed portion extending toward the interior of said container.
33. An automated laboratory as in claim 32 wherein said container comprises an upper rim and said closure means comprises an annular cavity portion surrounding said duck-billed portion for sealing to said upper rim.
34. A duck-billed closure for a container to permit easy entry of a needle-like device while minimizing exposure of a material inside said container, said closure comprising seal means for sealing to said container and a flexible duck-billed portion, said duck-billed portion extending toward the interior of said container with said closure sealed to said container.
35. A duck-billed closure as in claim 34 wherein said container comprises an upper rim and said closure means comprises an annular cavity portion surrounding said duck-billed portion for sealing to said upper rim.
36. A method for transferring liquid by robotic translation means from a first container holding a first volume of liquid to a second container holding a second volume of liquid comprising the steps of: aspirating a third volume of liquid from said first volume of liquid in said first container with a pipette means having a pipette tip and a sensing means for sensing the position of said pipette tip relative to proximate surfaces; moving said pipette tip away from said first container by action of said robotic translation means; dispensing a droplet of liquid from said pipette tip such that said droplet depends from said pipette tip but does not separate therefrom; moving said droplet with said pipette tip by said robotic translation means until said droplet touches the surface of said second volume of liquid, stopping translation when said sensing means signals contact, and allowing said droplet to become confluent with said second volume of liquid.
37. A method for aspirating a first volume of liquid from a second volume of liquid in a container comprising the steps of: moving a pipette means, said pipette means comprising a pipette tip and a position sensing means for sensing the position of said pipette tip relative to proximate surfaces, to the surface of said second volume of liquid by a robotic translation means; stopping translation of said pipette tip when said pipette tip touches said surface as signalled by said sensing means; and aspirating said first volume of liquid by said pipette means.
38. A method for aspirating a first volume of liquid as in claim 37 further comprising a step of moving said pipette tip downward while aspirating said first volume of liquid to track the surface of said second volume of liquid such that said pipette tip does not lose contact with said second volume of liquid while aspirating.
39. A method for aspirating a first volume of liquid as in claim 37 comprising a step of moving said pipette tip downward a fixed dimension to penetrate said surface after said step of moving said pipette tip to touch said surface and before said step of aspirating said first volume of liquid.
40. A method for validating a worksurface in a liquid-handling instrument having a pipette means, said pipette means comprising a pipette tip coupled to a position sensing means for sensing position of said pipette tip relative to proximate surfaces, said pipette tip movable over said worksurface by a robotic translation means, said method comprising the steps of: moving said pipette tip in a pattern and at a height over said worksurface such that said pipette tip will contact no surface if every part is in its proper place; stopping translation of said pipette tip if said pipette tip contacts any surface as signalled by said position sensing means; and activating a signal that a part is out of position.
41. A method for mixing liquids in a liquid handling instrument having a pipette means with a pipette tip movable over a worksurface by a robotic translation means, said method comprising the steps of: moving said pipette means by said robotic translation means to immerse said pipette tip in a volume of liquid in a container; aspirating liquid into said pipette means ending with said pipette tip immersed in said volume of liquid in said container; and dispensing said aspirated liquid into said container while moving said tip in a pattern encompassing substantially said volume of liquid in said container.
42. An automated laboratory as in claim 15 wherein said protocol comprises steps for performing automated specific gene detection.
43. An automated laboratory in claim 15 wherein said protocol comprises steps for performing automated nucleic acid sequence detection.
44. An automated laboratory in claim 15 wherein said protocol comprises steps for performing automated fluorescent labelling of nucleic acids.
PCT/US1991/002348 1990-04-06 1991-04-04 Automated molecular biology laboratory WO1991016675A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
DE69126690T DE69126690T2 (en) 1990-04-06 1991-04-04 AUTOMATED LABORATORY FOR MOLECULAR BIOLOGY
EP91908369A EP0478753B1 (en) 1990-04-06 1991-04-04 Automated molecular biology laboratory

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US50562690A 1990-04-06 1990-04-06
US505,826 1990-04-06

Publications (1)

Publication Number Publication Date
WO1991016675A1 true WO1991016675A1 (en) 1991-10-31

Family

ID=24011134

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1991/002348 WO1991016675A1 (en) 1990-04-06 1991-04-04 Automated molecular biology laboratory

Country Status (5)

Country Link
US (1) US5443791A (en)
EP (1) EP0478753B1 (en)
AT (1) ATE154981T1 (en)
DE (1) DE69126690T2 (en)
WO (1) WO1991016675A1 (en)

Cited By (54)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0505004A2 (en) * 1991-03-21 1992-09-23 Johnson & Johnson Clinical Diagnostics, Inc. Tip to surface spacing for optimum dispensing
WO1992022800A1 (en) * 1991-06-13 1992-12-23 Abbott Laboratories Liquid dispensing mechanism
EP0555739A1 (en) * 1992-02-13 1993-08-18 F. Hoffmann-La Roche Ag Automatic pipetting device
WO1993020612A2 (en) * 1992-04-02 1993-10-14 Baxter Deutschland Gmbh Automatic device for the photometric analysis of liquid samples
WO1993025912A2 (en) * 1992-06-09 1993-12-23 Medical Research Council Automated preparation of nucleic acids
WO1994008759A1 (en) * 1992-10-16 1994-04-28 Thomas Jefferson University Method and apparatus for robotically performing sanger dideoxynucleotide dna sequencing reactions
DE4416406A1 (en) * 1993-05-10 1994-11-17 Olympus Optical Co Output device and output method
EP0644426A1 (en) * 1993-09-17 1995-03-22 F. Hoffmann-La Roche Ag Analyser with a device for suspending particles, and suspension method therefor
US5410699A (en) * 1989-08-25 1995-04-25 International Business Machines Corp. Apparatus and method for loading BIOS from a diskette in a personal computer system
WO1995011083A2 (en) * 1993-10-22 1995-04-27 Abbott Laboratories Reaction tube and method of use to minimize contamination
EP0681160A1 (en) * 1994-05-02 1995-11-08 F. Hoffmann-La Roche AG Analyser with automatic checking of the straightness of a pipetting needle
EP0681184A1 (en) * 1994-05-02 1995-11-08 F. Hoffmann-La Roche AG Analyser with automatic determination of the reference position of a pipetting needle
US5496517A (en) * 1989-12-22 1996-03-05 Beckman Instruments, Inc. Laboratory workstation using thermal vaporization control
EP0707726A1 (en) * 1993-07-09 1996-04-24 Akzo Nobel N.V. Memory control device for an assay apparatus
WO1997015809A1 (en) * 1995-10-27 1997-05-01 Dynex Technologies (Guernsey) Ltd. Level sensor and washer unit
DE19626234A1 (en) * 1996-06-29 1998-01-02 Innova Gmbh Device for the contamination-free supply and removal of liquids
EP0899566A2 (en) * 1997-08-28 1999-03-03 Hitachi, Ltd. Analytical apparatus, liquid chromatography analyzer and a method therefor
EP0961118A2 (en) * 1998-05-25 1999-12-01 Basf Aktiengesellschaft Pipetting automat
US6060022A (en) * 1996-07-05 2000-05-09 Beckman Coulter, Inc. Automated sample processing system including automatic centrifuge device
WO2000056872A2 (en) * 1999-03-22 2000-09-28 Pangene Corporation High-throughput gene cloning and phenotypic screening
EP1081234A2 (en) * 1999-09-06 2001-03-07 Toyo Boseki Kabushiki Kaisha Apparatus for purifying nucleic acids and proteins
US6293750B1 (en) 1998-07-14 2001-09-25 Bayer Corporation Robotics for transporting containers and objects within an automated analytical instrument and service tool for servicing robotics
EP1164200A2 (en) * 2000-05-31 2001-12-19 Pfizer Products Inc. Method and device for drug-drug interaction testing sample preparation
WO2002005939A1 (en) * 2000-07-18 2002-01-24 Basf Aktiengesellschaft Method and device for carrying out the automated preparation and characterization of liquid multi-constituent systems
WO2002027035A2 (en) * 2000-09-28 2002-04-04 Pangene Corporation High-throughput gene cloning and phenotypic screening
EP1354185A1 (en) * 2001-01-24 2003-10-22 Gilson, Inc. Probe tip alignment for precision liquid handler
DE10241007A1 (en) * 2002-09-05 2004-03-18 Eppendorf Ag Distributing liquid samples from automated liquid stations to automated workstations, for use in molecular biology, comprises that original and target matrix layouts are established to give locations/target destinations
US6827478B2 (en) 2000-07-18 2004-12-07 Basf Aktiengesellschaft Method and device for carrying out the automated preparation and characterization of liquid multi-constituent systems
EP1881330A2 (en) * 2006-07-20 2008-01-23 Ortho-Clinical Diagnostics, Inc. Fluid metering in a metering zone
WO2008052758A1 (en) * 2006-10-31 2008-05-08 Bürkert Werke GmbH & Co. KG Modular laboratory apparatus for analysis and synthesis of liquids and method for analysis and synthesis of liquids
EP1947463A1 (en) * 2007-01-16 2008-07-23 Roche Diagnostics GmbH Collection of liquid analytical samples for clinical analytical purpose
EP1992952A3 (en) * 2007-05-15 2009-05-06 Hitachi High-Technologies Corporation Liquid dispensing apparatus
US7976794B2 (en) 2006-07-21 2011-07-12 Stratec Biomedical Systems Ag Positioning device for the positioning of pipettes
US8021495B2 (en) 2005-04-21 2011-09-20 Wako Pure Chemical Industries, Ltd. Pipette cleaning device and cleaning method
EP2535712A1 (en) * 2011-06-15 2012-12-19 F. Hoffmann-La Roche AG Analytical system for the preparation of biological material
EP2565260A2 (en) * 2010-04-30 2013-03-06 Bioneer Corporation Automatic biological sample purification device having a magnetic-field-applying unit, a method for extracting a target substance from a biological sample, and a protein expression and purification method
CN103111828A (en) * 2013-03-12 2013-05-22 苏州大学 Thin-wall structural member automatic arranging and positioning machine and arranging and positioning method before finish machining
US8546110B2 (en) 1998-05-01 2013-10-01 Gen-Probe Incorporated Method for detecting the presence of a nucleic acid in a sample
US8840848B2 (en) 2010-07-23 2014-09-23 Beckman Coulter, Inc. System and method including analytical units
US8973736B2 (en) 2011-11-07 2015-03-10 Beckman Coulter, Inc. Magnetic damping for specimen transport system
US9046506B2 (en) 2011-11-07 2015-06-02 Beckman Coulter, Inc. Specimen container detection
WO2012119118A3 (en) * 2011-03-03 2016-01-14 Life Technologies Corporation Sampling probes, systems, apparatuses, and methods
US9335336B2 (en) 2011-09-09 2016-05-10 Gen-Probe Incorporated Automated sample handling instrumentation, systems, processes, and methods
US9446418B2 (en) 2011-11-07 2016-09-20 Beckman Coulter, Inc. Robotic arm
US9482684B2 (en) 2011-11-07 2016-11-01 Beckman Coulter, Inc. Centrifuge system and workflow
US9506943B2 (en) 2011-11-07 2016-11-29 Beckman Coulter, Inc. Aliquotter system and workflow
EP3147669A1 (en) * 2015-09-25 2017-03-29 Kabushiki Kaisha Yaskawa Denki Method for controlling pipetting robot and pipetting robot system
US9910054B2 (en) 2011-11-07 2018-03-06 Beckman Coulter, Inc. System and method for processing samples
CN108593944A (en) * 2018-03-22 2018-09-28 广州市第人民医院(广州消化疾病中心、广州医科大学附属市人民医院、华南理工大学附属第二医院) Liquid drop chip immunoassay system and method
CN110305773A (en) * 2019-08-22 2019-10-08 深圳市芯思微生物科技有限公司 A kind of nucleic acid-extracting apparatus and extracting method
EP3563929A1 (en) * 2018-04-30 2019-11-06 Firmenich SA Apparatus for customized production of a flavoring agent mix
EP3620772A1 (en) * 2018-09-06 2020-03-11 Milestone S.r.l. System and method for filling a closed container with a fixative solution
EP3080616B1 (en) 2013-12-12 2020-06-03 Diagnostica Stago Method of determining the position of at least one cartography token
CN112191207A (en) * 2020-09-28 2021-01-08 苏州市启献智能科技有限公司 Chemical experiment system of Internet of things

Families Citing this family (335)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6569620B1 (en) * 1990-06-11 2003-05-27 Somalogic, Inc. Method for the automated generation of nucleic acid ligands
US6716580B2 (en) * 1990-06-11 2004-04-06 Somalogic, Inc. Method for the automated generation of nucleic acid ligands
US5762876A (en) * 1991-03-05 1998-06-09 Molecular Tool, Inc. Automatic genotype determination
US5897783A (en) * 1992-09-24 1999-04-27 Amersham International Plc Magnetic separation method
RU2041263C1 (en) * 1993-08-11 1995-08-09 Геннадий Моисеевич Ершов Method and apparatus for microdosing and dispensing of aqueous solutions onto carrier
US5525300A (en) * 1993-10-20 1996-06-11 Stratagene Thermal cycler including a temperature gradient block
JPH09507027A (en) 1993-12-23 1997-07-15 モレキュラー・ツール・インク Automatic genotyping
DE4412286A1 (en) * 1994-04-09 1995-10-12 Boehringer Mannheim Gmbh System for contamination-free processing of reaction processes
AU3096795A (en) * 1994-07-11 1996-02-09 Akzo Nobel N.V. Micro sample tube with reduced dead volume and bar code capability
US5508197A (en) * 1994-07-25 1996-04-16 The Regents, University Of California High-speed thermal cycling system and method of use
US5601980A (en) * 1994-09-23 1997-02-11 Hewlett-Packard Company Manufacturing method and apparatus for biological probe arrays using vision-assisted micropipetting
US5623592A (en) * 1994-10-18 1997-04-22 Molecular Dynamics Method and apparatus for constructing an iconic sequence to operate external devices
JP3481705B2 (en) * 1994-12-12 2003-12-22 株式会社モリテックス Automatic solid-phase extraction device
US6983227B1 (en) 1995-01-17 2006-01-03 Intertech Ventures, Ltd. Virtual models of complex systems
US5930154A (en) * 1995-01-17 1999-07-27 Intertech Ventures, Ltd. Computer-based system and methods for information storage, modeling and simulation of complex systems organized in discrete compartments in time and space
US5980096A (en) * 1995-01-17 1999-11-09 Intertech Ventures, Ltd. Computer-based system, methods and graphical interface for information storage, modeling and stimulation of complex systems
US5551487A (en) * 1995-03-10 1996-09-03 Hewlett-Packard Company Micro-dispenser for preparing assay plates
US6171555B1 (en) 1995-04-17 2001-01-09 Ontogen Corporation Reaction block docking station
US5609826A (en) * 1995-04-17 1997-03-11 Ontogen Corporation Methods and apparatus for the generation of chemical libraries
US5772962A (en) * 1995-05-29 1998-06-30 Hitachi, Ltd. Analyzing apparatus using disposable reaction vessels
KR100463475B1 (en) 1995-06-08 2005-06-22 로셰 디아그노스틱스 게엠베하 Magnetic Pigment
DE19520398B4 (en) * 1995-06-08 2009-04-16 Roche Diagnostics Gmbh Magnetic pigment
US5926387A (en) * 1995-06-30 1999-07-20 Beckman Instruments, Inc. Ultracentrifuge operation by computer system
US5790119A (en) * 1995-10-30 1998-08-04 Xerox Corporation Apparatus and method for programming a job ticket in a document processing system
US6063633A (en) * 1996-02-28 2000-05-16 The University Of Houston Catalyst testing process and apparatus
US6048691A (en) * 1996-05-13 2000-04-11 Motorola, Inc. Method and system for performing a binding assay
US6054099A (en) * 1996-05-15 2000-04-25 Levy; Abner Urine specimen container
US6175816B1 (en) * 1997-05-23 2001-01-16 Advanced Life Sciences, Inc. Use of automated technology in chemical process research and development
US6044212A (en) * 1996-05-24 2000-03-28 Advanced Life Sciences, Inc. Use of automated technology in chemical process research and development
ES2216080T3 (en) * 1996-05-29 2004-10-16 Walter Dr. Schubert AUTOMATED DEVICE AND PROCEDURE FOR MEASURING AND DETERMINING MOLECULES OR PARTS OF MOLECULES.
WO1998010265A1 (en) * 1996-09-09 1998-03-12 Tyco Group S.A.R.L. Electronically monitored mechanical pipette
EP0944739A4 (en) 1996-09-16 2000-01-05 Univ Utah Res Found Method and apparatus for analysis of chromatographic migration patterns
US5795784A (en) 1996-09-19 1998-08-18 Abbott Laboratories Method of performing a process for determining an item of interest in a sample
US5856194A (en) 1996-09-19 1999-01-05 Abbott Laboratories Method for determination of item of interest in a sample
DE19643721A1 (en) * 1996-10-23 1998-05-07 Deutsches Krebsforsch Automated quantification of DNA strand breaks in intact cells
US5736403A (en) * 1996-11-13 1998-04-07 Johnson & Johnson Clinical Diagnostics, Inc. Determining height variations around a rotor
US5753512A (en) * 1996-11-13 1998-05-19 Johnson & Johnson Clinical Diagnostics, Inc Determining liquid volumes in cup-like vessels on a rotor having vertical deviations
JP4611462B2 (en) * 1996-11-13 2011-01-12 オルソ−クリニカル ダイアグノスティクス,インコーポレイティド Method for measuring liquid volume, apparatus therefor and method for confirming vertical position of reaction vessel
US5985214A (en) 1997-05-16 1999-11-16 Aurora Biosciences Corporation Systems and methods for rapidly identifying useful chemicals in liquid samples
US6258326B1 (en) 1997-09-20 2001-07-10 Ljl Biosystems, Inc. Sample holders with reference fiducials
US5958789A (en) * 1997-07-29 1999-09-28 Johnson & Johnson Clinical Diagnostics, Inc. Reduction in positive bias in wet assays due to splashing
ES2222538T3 (en) 1997-08-08 2005-02-01 Aventis Pharma Deutschland Gmbh PIPETED ROBOT WITH THERMOSTATIC DEVICE.
US20050135972A1 (en) * 1997-08-11 2005-06-23 Ventana Medical Systems, Inc. Method and apparatus for modifying pressure within a fluid dispenser
US6045759A (en) * 1997-08-11 2000-04-04 Ventana Medical Systems Fluid dispenser
US8137619B2 (en) * 1997-08-11 2012-03-20 Ventana Medical Systems, Inc. Memory management method and apparatus for automated biological reaction system
US6093574A (en) 1997-08-11 2000-07-25 Ventana Medical Systems Method and apparatus for rinsing a microscope slide
US7745142B2 (en) 1997-09-15 2010-06-29 Molecular Devices Corporation Molecular modification assays
EP1027145A4 (en) * 1997-09-17 2004-08-25 Gentra Systems Inc Apparatuses and methods for isolating nucleic acid
WO2000066269A1 (en) * 1999-05-03 2000-11-09 Ljl Biosystems, Inc. Integrated sample-processing system
US6838051B2 (en) 1999-05-03 2005-01-04 Ljl Biosystems, Inc. Integrated sample-processing system
US6902703B2 (en) 1999-05-03 2005-06-07 Ljl Biosystems, Inc. Integrated sample-processing system
US6297018B1 (en) 1998-04-17 2001-10-02 Ljl Biosystems, Inc. Methods and apparatus for detecting nucleic acid polymorphisms
DE19743518A1 (en) * 1997-10-01 1999-04-15 Roche Diagnostics Gmbh Automated, universally applicable sample preparation method
JP4564165B2 (en) 1997-10-31 2010-10-20 アプライド バイオシステムズ, エルエルシー Method and apparatus for making array (s)
DE29720432U1 (en) 1997-11-19 1999-03-25 Mwg Biotech Gmbh robot
AUPP058197A0 (en) * 1997-11-27 1997-12-18 A.I. Scientific Pty Ltd Pathology sample tube distributor
US6893877B2 (en) * 1998-01-12 2005-05-17 Massachusetts Institute Of Technology Methods for screening substances in a microwell array
US6722395B2 (en) * 1998-01-13 2004-04-20 James W. Overbeck Depositing fluid specimens on substrates, resulting ordered arrays, techniques for analysis of deposited arrays
US6428752B1 (en) 1998-05-14 2002-08-06 Affymetrix, Inc. Cleaning deposit devices that form microarrays and the like
US6269846B1 (en) 1998-01-13 2001-08-07 Genetic Microsystems, Inc. Depositing fluid specimens on substrates, resulting ordered arrays, techniques for deposition of arrays
US6407858B1 (en) 1998-05-14 2002-06-18 Genetic Microsystems, Inc Focusing of microscopes and reading of microarrays
AU747296B2 (en) * 1998-02-10 2002-05-16 Toyo Kohan Co. Ltd. Apparatus for immobilized DNA library preparation, apparatus for gene amplification, method for temperature control and method for comparing genes systematically
US8192994B2 (en) 1998-02-10 2012-06-05 Angros Lee H Method of applying a biological specimen to an analytic plate
US6660228B1 (en) * 1998-03-02 2003-12-09 Cepheid Apparatus for performing heat-exchanging, chemical reactions
US7095032B2 (en) * 1998-03-20 2006-08-22 Montagu Jean I Focusing of microscopes and reading of microarrays
US8337753B2 (en) 1998-05-01 2012-12-25 Gen-Probe Incorporated Temperature-controlled incubator having a receptacle mixing mechanism
US6267020B1 (en) * 1998-07-30 2001-07-31 Universal Instruments Corporation Drive mechanism for variable center distance component insertion machine
EP0977037B1 (en) * 1998-07-31 2005-08-31 Tecan Trading AG Magnetic separator
US6759014B2 (en) * 2001-01-26 2004-07-06 Symyx Technologies, Inc. Apparatus and methods for parallel processing of multiple reaction mixtures
US6913934B2 (en) * 1998-08-13 2005-07-05 Symyx Technologies, Inc. Apparatus and methods for parallel processing of multiple reaction mixtures
US6890492B1 (en) 1998-08-13 2005-05-10 Symyx Technologies, Inc. Parallel reactor with internal sensing and method of using same
US6548026B1 (en) 1998-08-13 2003-04-15 Symyx Technologies, Inc. Parallel reactor with internal sensing and method of using same
US6528026B2 (en) 1998-08-13 2003-03-04 Symyx Technologies, Inc. Multi-temperature modular reactor and method of using same
US6455316B1 (en) 1998-08-13 2002-09-24 Symyx Technologies, Inc. Parallel reactor with internal sensing and method of using same
US6306658B1 (en) 1998-08-13 2001-10-23 Symyx Technologies Parallel reactor with internal sensing
US6270249B1 (en) 1998-09-30 2001-08-07 Robert W. Besuner Vertically reciprocating perforated agitator
US6413780B1 (en) * 1998-10-14 2002-07-02 Abbott Laboratories Structure and method for performing a determination of an item of interest in a sample
CA2346268A1 (en) * 1998-10-16 2000-04-27 Intelligent Automation Systems Continuous processing automated workstation
US7199809B1 (en) * 1998-10-19 2007-04-03 Symyx Technologies, Inc. Graphic design of combinatorial material libraries
US6592820B1 (en) * 1998-11-05 2003-07-15 Bio-Spectrum Technologies, Inc. System and method for biochemical assay
DE69924645T2 (en) * 1998-11-13 2006-03-02 Cellomics, Inc. METHOD AND SYSTEM FOR EFFICIENTLY GAINING AND STORING EXPERIMENTAL DATA
DE19859586C1 (en) * 1998-12-22 2000-07-13 Mwg Biotech Ag Thermal cycler device
DE29925002U1 (en) 1998-12-22 2008-06-19 Applera Corp. (n.d.Ges.d. Staates Delaware), Foster City thermocycler
US6443022B1 (en) 1999-01-14 2002-09-03 Intelligent Automation Systems, Inc. Fluid level detector and method for use with automated workstation
US20030054360A1 (en) * 1999-01-19 2003-03-20 Larry Gold Method and apparatus for the automated generation of nucleic acid ligands
MXPA01007352A (en) * 1999-01-19 2003-06-06 Somalogic Inc Method and apparatus for the automated generation of nucleic acid ligands.
US6270644B1 (en) 1999-01-27 2001-08-07 Affymetrix, Inc. Capillary array electrophoresis scanner
DE60030957T2 (en) * 1999-02-16 2007-06-14 Applera Corp., Foster City BALL DELIVERY SYSTEM
US6690168B1 (en) * 1999-02-22 2004-02-10 Allen Ray Herron Biomagnetic detecting and imaging device
US6296702B1 (en) 1999-03-15 2001-10-02 Pe Corporation (Ny) Apparatus and method for spotting a substrate
US20020176801A1 (en) * 1999-03-23 2002-11-28 Giebeler Robert H. Fluid delivery and analysis systems
US6296771B1 (en) 1999-04-02 2001-10-02 Symyx Technologies, Inc. Parallel high-performance liquid chromatography with serial injection
US6436292B1 (en) * 1999-04-02 2002-08-20 Symyx Technologies, Inc. Parallel high-performance liquid chromatography with post-separation treatment
AU4226900A (en) * 1999-04-08 2000-10-23 Joseph L. Chan Apparatus for fast preparation and analysis of nucleic acids
US6245297B1 (en) * 1999-04-16 2001-06-12 Pe Corporation (Ny) Apparatus and method for transferring small volumes of substances
DE19918956A1 (en) * 1999-04-27 2000-11-02 Studiengesellschaft Kohle Mbh Process for the automated investigation of catalytic and spectroscopic properties of the components of combinatorial libraries
US6507945B1 (en) 1999-05-05 2003-01-14 Symyx Technologies, Inc. Synthesizing combinatorial libraries of materials
US6716396B1 (en) 1999-05-14 2004-04-06 Gen-Probe Incorporated Penetrable cap
CA2678141C (en) 1999-05-14 2011-09-06 Gen-Probe Incorporated Collection device comprising penetrable cap and method for use thereof
US6573369B2 (en) * 1999-05-21 2003-06-03 Bioforce Nanosciences, Inc. Method and apparatus for solid state molecular analysis
US20030186311A1 (en) * 1999-05-21 2003-10-02 Bioforce Nanosciences, Inc. Parallel analysis of molecular interactions
US20030073250A1 (en) * 1999-05-21 2003-04-17 Eric Henderson Method and apparatus for solid state molecular analysis
US20020042081A1 (en) * 2000-10-10 2002-04-11 Eric Henderson Evaluating binding affinities by force stratification and force panning
WO2000072969A1 (en) * 1999-05-27 2000-12-07 The Perkin-Elmer Corporation Apparatus and method for the precise location of reaction plates
AU5878900A (en) * 1999-06-19 2001-01-09 Orchid Biosciences, Inc. Microfluidic device interface
US6508986B1 (en) * 1999-08-27 2003-01-21 Large Scale Proteomics Corp. Devices for use in MALDI mass spectrometry
US7167615B1 (en) 1999-11-05 2007-01-23 Board Of Regents, The University Of Texas System Resonant waveguide-grating filters and sensors and methods for making and using same
EP1099950A1 (en) * 1999-11-12 2001-05-16 F. Hoffmann-La Roche Ag Analyzer having a rotatable sample rack carrier
PL202358B1 (en) * 1999-11-17 2009-06-30 Roche Diagnostics Gmbh Magnetic glass particles, method for their preparation and uses thereof
DE60030882T2 (en) * 2000-01-06 2007-04-05 Caliper Life Sciences, Inc., Mountain View DEVICES AND METHODS FOR HIGH-SPEED SAMPLE TAKING AND ANALYSIS
US20030097222A1 (en) * 2000-01-25 2003-05-22 Craford David M. Method, system, and computer software for providing a genomic web portal
EP1252598A2 (en) 2000-01-25 2002-10-30 Cellomics, Inc. Method and system for automated inference of physico-chemical interaction knowledge
US6325114B1 (en) 2000-02-01 2001-12-04 Incyte Genomics, Inc. Pipetting station apparatus
US6932895B2 (en) * 2000-02-15 2005-08-23 Large Scale Proteomics Corporation Automated electrophoresis gel manipulation apparatus
DE10006846C2 (en) * 2000-02-16 2002-03-07 Macherey Nagel Gmbh & Co Hg Process for photometric COD measurement
CA2400644C (en) * 2000-02-18 2009-07-14 Board Of Trustees Of The Leland Stanford Junior University Apparatus and methods for parallel processing of micro-volume liquid reactions
JP4717312B2 (en) 2000-02-29 2011-07-06 ジェン−プローブ・インコーポレイテッド Fluid transfer probe
US6897015B2 (en) * 2000-03-07 2005-05-24 Bioforce Nanosciences, Inc. Device and method of use for detection and characterization of pathogens and biological materials
EP1186881A4 (en) * 2000-03-16 2006-04-19 Fuji Photo Film Co Ltd Measuring method and instrument utilizing total reflection attenuation
SE0001196D0 (en) * 2000-04-03 2000-04-03 Alfa Laval Agri Ab Milk sampling apparatus and method
SE517143C2 (en) * 2000-04-03 2002-04-23 Delaval Holding Ab Device and method for sampling milk
DE20006548U1 (en) * 2000-04-08 2001-08-23 Mwg Biotech Ag Device for carrying out chemical or biological processes
DE10017790A1 (en) * 2000-04-10 2001-10-11 Basf Ag Process for the production of biopolymer fields with real-time control
US6672458B2 (en) * 2000-05-19 2004-01-06 Becton, Dickinson And Company System and method for manipulating magnetically responsive particles fluid samples to collect DNA or RNA from a sample
US6994827B2 (en) 2000-06-03 2006-02-07 Symyx Technologies, Inc. Parallel semicontinuous or continuous reactors
AU2001268481A1 (en) * 2000-06-15 2001-12-24 Board Of Regents, The University Of Texas System Regulatable, catalytically active nucleic acids
JP2004503782A (en) * 2000-06-15 2004-02-05 アイアールエム,エルエルシー Automatic precision object holder
JP2002001092A (en) * 2000-06-22 2002-01-08 Shimadzu Corp Apparatus for discharging liquid
JP2002022752A (en) * 2000-07-13 2002-01-23 Suzuki Motor Corp Specimen testing device
US7018589B1 (en) 2000-07-19 2006-03-28 Symyx Technologies, Inc. High pressure parallel reactor
US7008769B2 (en) * 2000-08-15 2006-03-07 Bioforce Nanosciences, Inc. Nanoscale molecular arrayer
US6789040B2 (en) 2000-08-22 2004-09-07 Affymetrix, Inc. System, method, and computer software product for specifying a scanning area of a substrate
US6768982B1 (en) 2000-09-06 2004-07-27 Cellomics, Inc. Method and system for creating and using knowledge patterns
CA2423552A1 (en) * 2000-10-13 2002-04-18 Irm Llc High throughput processing system and method of using
US7714301B2 (en) * 2000-10-27 2010-05-11 Molecular Devices, Inc. Instrument excitation source and calibration method
AU2002249778A1 (en) 2000-11-17 2002-08-12 Thermogenic Imaging, Inc. Apparatus and methods for infrared calorimetric measurements
US20020132360A1 (en) 2000-11-17 2002-09-19 Flir Systems Boston, Inc. Apparatus and methods for infrared calorimetric measurements
US7435390B2 (en) * 2001-01-26 2008-10-14 Third Wave Technologies, Inc. Nucleic acid synthesizers
US20030072689A1 (en) * 2001-08-15 2003-04-17 Third Wave Technologies, Inc. Polymer synthesizer
US20080261220A1 (en) * 2000-11-30 2008-10-23 Third Wave Technologies, Inc. Nucleic Acid Detection Assays
US6932943B1 (en) 2001-01-26 2005-08-23 Third Wave Technologies Nucleic acid synthesizers
EP1354211A1 (en) * 2001-01-25 2003-10-22 Tecan Trading AG Pipetting device
FR2820756B1 (en) * 2001-02-09 2004-01-23 Daniel Attias INCUBATOR AND INCUBATION PROCESS ENDING THE ORGANIZATION SET TO INCUBATE
US6692700B2 (en) 2001-02-14 2004-02-17 Handylab, Inc. Heat-reduction methods and systems related to microfluidic devices
EP1370876A2 (en) * 2001-02-16 2003-12-17 Aventis Pharmaceuticals Inc. Automated semi-solid matrix assay and liquid handler apparatus for the same
GB0117706D0 (en) * 2001-02-16 2001-09-12 Aventis Pharm Prod Inc Automated semi-solid matrix assay and liquid handler apparatus for the same
US6852287B2 (en) 2001-09-12 2005-02-08 Handylab, Inc. Microfluidic devices having a reduced number of input and output connections
US7010391B2 (en) 2001-03-28 2006-03-07 Handylab, Inc. Methods and systems for control of microfluidic devices
US7829025B2 (en) 2001-03-28 2010-11-09 Venture Lending & Leasing Iv, Inc. Systems and methods for thermal actuation of microfluidic devices
US8895311B1 (en) 2001-03-28 2014-11-25 Handylab, Inc. Methods and systems for control of general purpose microfluidic devices
EP1251345A1 (en) * 2001-04-12 2002-10-23 Fuji Photo Film Co., Ltd. Measuring sensor utilizing attenuated total reflection and measuring chip assembly
EP1252930A1 (en) * 2001-04-27 2002-10-30 Tecan Trading AG Spray head
US6790413B2 (en) 2001-05-03 2004-09-14 Beckman Coulter, Inc. Sample presentation unit
US20020168292A1 (en) * 2001-05-14 2002-11-14 Whisenhunt Donald Wayne Systems and methods for the high throughput preparation and analysis of chemical reactions
US6485918B1 (en) 2001-07-02 2002-11-26 Packard Bioscience Corporation Method and apparatus for incubation of a liquid reagent and target spots on a microarray substrate
WO2003008941A2 (en) * 2001-07-17 2003-01-30 Bioforce Nanosciences, Inc. Combined molecular blinding detection through force microscopy and mass spectrometry
EP1291659A3 (en) * 2001-09-06 2008-05-21 Sysmex Corporation Automatic sample analyzer and its components
US6579724B2 (en) * 2001-09-13 2003-06-17 First Ten Angstroms Dispensing method and apparatus for dispensing very small quantities of fluid
US7042488B2 (en) 2001-09-27 2006-05-09 Fujinon Corporation Electronic endoscope for highlighting blood vessel
JP2003149255A (en) * 2001-11-15 2003-05-21 Toyobo Co Ltd Dispensing device movable to rectangular coordinate and cyclinderical coordinate
US7036653B2 (en) 2002-01-29 2006-05-02 Siemens Technology-To-Business Center Llc Load manipulation system
US6910569B2 (en) 2002-01-29 2005-06-28 Siemens Technology-To-Business Center, Llc Load singulation system and method
US7378058B2 (en) * 2002-01-30 2008-05-27 Ventana Medical Systems, Inc. Method and apparatus for modifying pressure within a fluid dispenser
US20050118067A1 (en) * 2002-02-12 2005-06-02 Jaan Noolandi Device to print biofluids
DE10212557A1 (en) * 2002-03-14 2003-09-25 Univ Schiller Jena Characterizing large numbers of samples with volume in microliter or sub-microliter range in parallel uses solutions containing two different indicators to produce signals of different intensities
US6937955B2 (en) * 2002-03-29 2005-08-30 Ortho-Clinical Diagnostics, Inc. Method for automatic alignment of metering system for a clinical analyzer
EP1499187B1 (en) 2002-04-04 2015-06-17 Zoetis Belgium S.A. Immunostimulatory g,u-containing oligoribonucleotides
US7361509B2 (en) * 2002-04-29 2008-04-22 Ortho-Clinical Diagnostics Dynamic metered fluid volume determination method and related apparatus
US7122159B2 (en) * 2002-04-29 2006-10-17 Symyx Technologies, Inc. High pressure parallel reactor with individually sealable vessels
MXPA04011083A (en) * 2002-05-17 2005-02-14 Becton Dickinson Co Automated system for isolating, amplyifying and detecting a target nucleic acid sequence.
US20030213906A1 (en) * 2002-05-20 2003-11-20 Large Scale Proteomics Corporation Method and apparatus for minimizing evaporation of a volatile substance
US20050239193A1 (en) * 2002-05-30 2005-10-27 Bioforce Nanosciences, Inc. Device and method of use for detection and characterization of microorganisms and microparticles
WO2004011680A1 (en) * 2002-07-25 2004-02-05 Archemix Corp. Regulated aptamer therapeutics
US20040018635A1 (en) * 2002-07-26 2004-01-29 Peck Bill J. Fabricating arrays with drop velocity control
US20060051493A1 (en) * 2002-07-31 2006-03-09 Tella Richard P Apparatus and methods for printing arrays
US8277753B2 (en) 2002-08-23 2012-10-02 Life Technologies Corporation Microfluidic transfer pin
US7261800B1 (en) * 2002-08-26 2007-08-28 Helena Laboratories Automatic in situ electrophoresis method and apparatus
EP1548437A4 (en) * 2002-09-17 2006-06-07 Olympus Corp Method and apparatus for arranging liquid reaction components on substrate surface for detecting target substance by reaction among plural reaction components on substrate and article utilized in the method
AU2003270568A1 (en) * 2002-09-17 2004-04-08 Pharmacia Corporation Preservation of rna and reverse transcriptase during automated liquid handling
EP1556297B1 (en) * 2002-10-29 2008-04-23 Siemens Aktiengesellschaft Conveyor system with distributed article manipulation
JP4045928B2 (en) * 2002-11-15 2008-02-13 日立工機株式会社 Automatic dispensing device
AU2003278682B2 (en) * 2002-11-18 2007-07-19 Agency For Science, Technology And Research Method and system for cell and/or nucleic acid molecules isolation
JP3985665B2 (en) * 2002-11-18 2007-10-03 日立工機株式会社 Automatic dispensing device
DE10253939A1 (en) * 2002-11-19 2004-06-17 Cetoni Gmbh Innovative Informationssysteme Process for the exact three-dimensional micropositioning of bodies of different geometry and / or dimensions in space
CA2521999A1 (en) 2002-12-20 2004-09-02 Biotrove, Inc. Assay apparatus and method using microfluidic arrays
US7344832B2 (en) * 2003-01-02 2008-03-18 Bioforce Nanosciences, Inc. Method and apparatus for molecular analysis in small sample volumes
JP3711988B2 (en) * 2003-05-12 2005-11-02 株式会社日立製作所 Fine particle array analysis system, fine particle array kit, and chemical analysis method
US7186378B2 (en) * 2003-07-18 2007-03-06 Dade Behring Inc. Liquid sampling probe and cleaning fluidics system
US20050014284A1 (en) * 2003-07-18 2005-01-20 Merrit Jacobs Improved fluid mixing in a diagnostic analyzer
US7731906B2 (en) 2003-07-31 2010-06-08 Handylab, Inc. Processing particle-containing samples
JP4251627B2 (en) * 2003-09-19 2009-04-08 株式会社東芝 Chemical analyzer and dispensing method thereof
DE10346286B3 (en) 2003-10-06 2005-04-14 J. Eberspächer GmbH & Co. KG The exhaust purification device
US20050065633A1 (en) * 2003-11-14 2005-03-24 Michael Wynblatt Systems and methods for relative control of load motion actuators
US20050107911A1 (en) * 2003-11-14 2005-05-19 Siemens Technology-To-Business Center Llc Systems and methods for controlling load motion actuators
US20050107909A1 (en) * 2003-11-14 2005-05-19 Siemens Technology-To-Business Center Llc Systems and methods for programming motion control
ATE320009T1 (en) * 2004-01-15 2006-03-15 Agilent Technologies Inc POSITIONING SYSTEM AND METHOD FOR A LIQUID TRANSFER DEVICE
JP4542345B2 (en) * 2004-01-15 2010-09-15 シスメックス株式会社 Analysis equipment
US20050164375A1 (en) * 2004-01-23 2005-07-28 Sysmex Corporation Nucleic acid detection apparatus
JP2007529015A (en) 2004-03-12 2007-10-18 バイオトローブ, インコーポレイテッド Nanoliter array loading
JP2007535681A (en) * 2004-04-30 2007-12-06 バイオフォース・ナノサイエンシィズ・インコーポレーテッド Method and apparatus for depositing material on a surface
US8852862B2 (en) 2004-05-03 2014-10-07 Handylab, Inc. Method for processing polynucleotide-containing samples
US20070244314A1 (en) * 2004-05-18 2007-10-18 Fujifilm Corporation Method For Extracting Nucleic Acid And Nucleic Acid-Extracting Apparatus
US7222526B2 (en) 2004-06-17 2007-05-29 Ortho-Clinical Diagnostics, Inc Liquid measurements using capacitive monitoring
JP2006090718A (en) * 2004-09-21 2006-04-06 Fuji Photo Film Co Ltd Sensor in utilizing attenuated total reflection
ATE390562T1 (en) * 2004-11-19 2008-04-15 Ebm Papst St Georgen Gmbh & Co ARRANGEMENT WITH A FAN AND A PUMP
US7258480B2 (en) * 2005-01-10 2007-08-21 Dade Behring Inc. Apparatus for mixing liquid samples using a two dimensional stirring pattern
JP2008532526A (en) 2005-03-10 2008-08-21 ジェン−プロウブ インコーポレイテッド System and method for performing an assay for detecting or quantifying an analyte in a sample
US20060246576A1 (en) * 2005-04-06 2006-11-02 Affymetrix, Inc. Fluidic system and method for processing biological microarrays in personal instrumentation
US7731811B2 (en) * 2005-04-15 2010-06-08 Angros Lee H Analytic substrate coating apparatus and method
US7968096B2 (en) * 2005-05-04 2011-06-28 The University Of Vermont And State Agricultural College Methods and compositions for treating toxoplasma
JP2006343299A (en) * 2005-05-12 2006-12-21 Uniflows Co Ltd Liquid feeder
JP5139976B2 (en) 2005-05-16 2013-02-06 インテリジェント ホスピタル システムズ リミテッド Automatic pharmacy mixing system (APAS)
US7534081B2 (en) * 2005-05-24 2009-05-19 Festo Corporation Apparatus and method for transferring samples from a source to a target
US8245740B2 (en) * 2005-06-03 2012-08-21 Alfa Wassermann, Inc. Fraction collector
DE202005014704U1 (en) * 2005-09-16 2007-02-01 C. Gerhardt Fabrik Und Lager Chemischer Apparate Gmbh & Co. Kg Device for the preparation of oil compositions for aromatherapy
ES2313200T3 (en) * 2005-09-21 2009-03-01 F. Hoffmann-La Roche Ag METHOD AND APPLIANCE FOR THE ACCURATE POSITIONING OF A PIPETING DEVICE.
US20070092403A1 (en) * 2005-10-21 2007-04-26 Alan Wirbisky Compact apparatus, compositions and methods for purifying nucleic acids
US20100072272A1 (en) * 2005-10-26 2010-03-25 Angros Lee H Microscope slide coverslip and uses thereof
US20100073766A1 (en) * 2005-10-26 2010-03-25 Angros Lee H Microscope slide testing and identification assembly
DE102006003117A1 (en) * 2006-01-18 2007-07-19 Institut für Bioprozess- und Analysenmesstechnik e.V. Energy transferring device for use in e.g. mini environment box, has inner carriage and rotors converted into movement having more than degree of freedom or into two movements having just one degree of freedom
EP1979751A1 (en) * 2006-01-25 2008-10-15 Koninklijke Philips Electronics N.V. Device for analyzing fluids
AU2007211847B2 (en) * 2006-02-02 2011-01-27 Qiagen Instruments Ag Thermocycler and sample port
JP5219181B2 (en) * 2006-02-02 2013-06-26 武蔵エンジニアリング株式会社 Pallet for fixing workpiece and liquid application apparatus having the same
US7998708B2 (en) 2006-03-24 2011-08-16 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US10900066B2 (en) 2006-03-24 2021-01-26 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
US11806718B2 (en) 2006-03-24 2023-11-07 Handylab, Inc. Fluorescence detector for microfluidic diagnostic system
WO2007112114A2 (en) 2006-03-24 2007-10-04 Handylab, Inc. Integrated system for processing microfluidic samples, and method of using same
EP2013662B1 (en) 2006-04-19 2013-08-14 Northwestern University Article for parallel lithography with two-dimensional pen arrays
US8192794B2 (en) * 2006-04-19 2012-06-05 Northwestern University Massively parallel lithography with two-dimensional pen arrays
KR20090049578A (en) 2006-06-28 2009-05-18 노쓰웨스턴유니버시티 Etching and hole arrays
CN1912627B (en) * 2006-08-23 2011-05-11 陕西北美基因股份有限公司 Blood treatment working station based on micronano magnetic particle and its control method
US20080056952A1 (en) * 2006-08-25 2008-03-06 Angros Lee H Analytic plates with markable portions and methods of use
US7901624B2 (en) 2006-09-26 2011-03-08 Becton, Dickinson And Company Device for automatically adjusting the bacterial inoculum level of a sample
WO2008060604A2 (en) 2006-11-14 2008-05-22 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
EP2091647A2 (en) 2006-11-14 2009-08-26 Handylab, Inc. Microfluidic system for amplifying and detecting polynucleotides in parallel
TWI388831B (en) * 2007-01-30 2013-03-11 Academia Sinica In-solution microarray assay
JP5202339B2 (en) * 2007-02-07 2013-06-05 ユニバーサル・バイオ・リサーチ株式会社 Container repeated use magnetic particle parallel processing apparatus and container repeated use magnetic particle parallel processing method
JP2010521325A (en) * 2007-03-13 2010-06-24 ナノインク インコーポレーティッド Nanolithography using viewport
US7807109B2 (en) 2007-05-14 2010-10-05 Freeslate, Inc. Parallel batch reactor with pressure monitoring
US7655191B2 (en) * 2007-05-14 2010-02-02 Symyx Solutions, Inc. Methods for chemical reactions in a parallel batch reactor
US20080286170A1 (en) * 2007-05-14 2008-11-20 Symyx Technologies, Inc. Parallel batch reactor
US20120164396A1 (en) 2007-06-20 2012-06-28 Northwestern University Matrix assisted ink transport
US20090004063A1 (en) * 2007-06-29 2009-01-01 Symyx Technologies, Inc. Apparatus and method for actuating a syringe
US9186677B2 (en) 2007-07-13 2015-11-17 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US8133671B2 (en) 2007-07-13 2012-03-13 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
US8182763B2 (en) 2007-07-13 2012-05-22 Handylab, Inc. Rack for sample tubes and reagent holders
AU2013205253B8 (en) * 2007-07-13 2015-10-22 Handylab, Inc. Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
EP3741869A1 (en) 2007-07-13 2020-11-25 Handylab, Inc. Polynucleotide capture materials and methods of using same
US8287820B2 (en) 2007-07-13 2012-10-16 Handylab, Inc. Automated pipetting apparatus having a combined liquid pump and pipette head system
US8105783B2 (en) 2007-07-13 2012-01-31 Handylab, Inc. Microfluidic cartridge
US9618139B2 (en) 2007-07-13 2017-04-11 Handylab, Inc. Integrated heater and magnetic separator
JP2010536033A (en) * 2007-08-08 2010-11-25 ノースウエスタン ユニバーシティ Independently addressable self-correcting inking method for cantilever arrays
AU2008329813A1 (en) * 2007-11-26 2009-06-04 Nanoink, Inc. Cantilever with pivoting actuation
US8726745B2 (en) * 2007-11-26 2014-05-20 Perkinelmer Health Sciences, Inc. Fluid handling device with ultrasound sensor and methods and systems using same
CA2712430A1 (en) * 2008-01-25 2009-07-30 Luminex Corporation Assay preparation plates, fluid assay preparation and analysis systems, and methods for preparing and analyzing assays
EP2250533A2 (en) * 2008-02-05 2010-11-17 Nanoink, Inc. Array and cantilever array leveling
EP2613155B1 (en) * 2008-04-24 2014-04-30 Tecan Trading AG Direct pipetting in computer-controlled liquid handling workstations
US8473085B2 (en) * 2008-04-30 2013-06-25 Perkinelmer Las, Inc. Mutex-mediated control of spatial access by appliances moveable over a common physical space
WO2009140441A2 (en) * 2008-05-13 2009-11-19 Nanoink, Inc. Height sensing cantilever
US20090314861A1 (en) * 2008-06-18 2009-12-24 Jaan Noolandi Fluid ejection using multiple voltage pulses and removable modules
USD787087S1 (en) 2008-07-14 2017-05-16 Handylab, Inc. Housing
US7804599B2 (en) * 2008-07-24 2010-09-28 MGM Instruments, Inc. Fluid volume verification system
EP2350673B1 (en) 2008-10-24 2022-05-11 Leica Biosystems Richmond, Inc. Modular system for performing laboratory protocols and associated methods
US9103782B2 (en) 2008-12-02 2015-08-11 Malvern Instruments Incorporated Automatic isothermal titration microcalorimeter apparatus and method of use
ES2835181T3 (en) 2009-04-27 2021-06-22 El Spectra Llc Pipette instrument
BRPI1012879B1 (en) * 2009-05-15 2019-10-29 Bio Merieux Inc system and method for automatic ventilation and sampling of a culture specimen container
CN102460183B (en) 2009-05-15 2015-04-15 生物梅里埃有限公司 Automated loading mechanism for microbial detection apparatus
US8347925B2 (en) * 2009-08-19 2013-01-08 Institute Of Nuclear Energy Research Automatic filling machine for radiopharmaceuticals
TW201113523A (en) * 2009-08-31 2011-04-16 Mbio Diagnostics Inc Integrated sample preparation and analyte detection
JP4919119B2 (en) * 2010-01-19 2012-04-18 株式会社日立プラントテクノロジー Separation / dispensing method with reagent dispensing nozzle and reagent dispensing / dispensing mechanism
US10697990B2 (en) 2010-04-29 2020-06-30 Leica Biosystems Richmond, Inc. Analytical system for performing laboratory protocols and associated methods
WO2012006084A2 (en) 2010-06-28 2012-01-12 Life Technologies Corporation Systems and methods for transfer of liquid samples
US9046507B2 (en) 2010-07-29 2015-06-02 Gen-Probe Incorporated Method, system and apparatus for incorporating capacitive proximity sensing in an automated fluid transfer procedure
US20120034703A1 (en) * 2010-08-06 2012-02-09 Affymetrix, Inc. Devices, Systems and Methods for Processing of Magnetic Particles
WO2012033765A1 (en) * 2010-09-07 2012-03-15 The Arizona Board Of Regents On Behalf Of The University Of Arizona Microdroplet-manipulation systems and methods for automated execution of molecular biological protocols
EP2654955B1 (en) * 2010-12-20 2015-07-15 Boehringer Ingelheim Microparts GmbH Method for mixing at least one sample solution with reagents
WO2012116308A1 (en) 2011-02-24 2012-08-30 Gen-Probe Incorporated Systems and methods for distinguishing optical signals of different modulation frequencies in an optical signal detector
CN103597358B (en) 2011-04-15 2016-08-17 贝克顿·迪金森公司 Scan real-time microfluid thermal cycler and for the thermal cycle synchronized and the method for scanning optical detection
CN103501909A (en) * 2011-05-13 2014-01-08 艾克特瑞斯有限责任公司 Methods and systems for automated pipette tracking
JP5714410B2 (en) * 2011-05-16 2015-05-07 株式会社日立ハイテクノロジーズ Automatic analyzer and method
JP5815129B2 (en) 2011-06-01 2015-11-17 ベンタナ メディカル システムズ, インコーポレイテッド Dispenser with filter device
PT2535034E (en) * 2011-06-17 2014-07-28 Kiro Robotics Sl Machine and method for the automatic preparation of intravenous medication
CN105699677A (en) 2011-07-01 2016-06-22 贝克曼考尔特公司 Low carryover liquid handling probe for an automated analyzer
WO2013009654A1 (en) * 2011-07-08 2013-01-17 Life Technologies Corporation Method and apparatus for automated sample manipulation
CA2849917C (en) 2011-09-30 2020-03-31 Becton, Dickinson And Company Unitized reagent strip
USD692162S1 (en) 2011-09-30 2013-10-22 Becton, Dickinson And Company Single piece reagent holder
CN104040238B (en) 2011-11-04 2017-06-27 汉迪拉布公司 Polynucleotides sample preparation apparatus
AU2013214849B2 (en) 2012-02-03 2016-09-01 Becton, Dickinson And Company External files for distribution of molecular diagnostic tests and determination of compatibility between tests
US9649639B2 (en) * 2012-02-03 2017-05-16 Corning Incorporated Separation apparatus and methods of separating magnetic material
KR101762295B1 (en) * 2012-02-10 2017-08-04 (주)바이오니아 Automatic analysis apparatus and method of biological samples
JP6138173B2 (en) 2012-03-09 2017-05-31 ライカ バイオシステムズ リッチモンド インコーポレイテッドLeica Biosystems Richmond, Inc. Method and apparatus for controlling temperature in a dynamic fluid in a laboratory sample processing system
CN102954954B (en) * 2012-09-19 2014-07-16 东南大学 Magnetic separation-based multi-sample multi-site high-flux nucleic acid analysis system
FR2999012B1 (en) * 2012-11-30 2017-12-15 Primadiag S A S MAGNETIC ATTRACTION MODULE, ROBOT COMPRISING SUCH A MODULE, AND METHOD FOR USE ON MAGNETIC BALLS OF SUCH A MODULE OR ROBOT
JP2014119387A (en) * 2012-12-18 2014-06-30 Sony Corp Dispensing apparatus, analyzer and method for controlling dispensing apparatus
DE102013200193A1 (en) 2013-01-09 2014-07-10 Hamilton Bonaduz Ag Sample processing system with dosing device and thermocycler
WO2014144640A1 (en) 2013-03-15 2014-09-18 Abbott Laboratories Automated diagnostic analyzers having vertically arranged carousels and related methods
ES2901756T3 (en) 2013-03-15 2022-03-23 Abbott Lab Automated diagnostic analyzers having rear accessible track systems and related methods
CN108196079B (en) 2013-03-15 2021-10-08 雅培制药有限公司 Diagnostic analyzer with pre-processing carousel and related methods
GB2514178B (en) * 2013-05-17 2015-09-09 Proxeon Biosystems As Automated system for handling components of a chromatographic system
EP3057708A4 (en) 2013-10-15 2017-06-21 Bio Molecular Systems Pty Ltd Improved thermocycler
JP5928435B2 (en) * 2013-11-01 2016-06-01 株式会社安川電機 Robot system, inspection method, and inspection object production method
EP2982439B1 (en) * 2014-08-06 2017-10-11 Yantai AusBio Laboratories Co., Ltd. Reagent carrier unit with coupling section to permit pipettor arm attachment and handling
EP3256256B1 (en) 2015-02-13 2022-12-21 Siemens Healthcare Diagnostics Inc. Pipette cleaning methods and apparatus, neutralizing liquid vessels, and methods of reducing carryover
CN204613224U (en) * 2015-03-13 2015-09-02 欧蒙医学诊断(中国)有限公司 Aspirating needle and comprise the film bar automatic pilot of this aspirating needle
CN108027379B (en) 2015-06-26 2021-07-23 雅培实验室 Reaction vessel exchange device for diagnostic analysis apparatus
CN108027280B (en) 2015-06-26 2021-07-06 雅培实验室 Reaction vessel movement component for moving a reaction vessel from a processing track to a rotation device in a diagnostic analyzer
GB201601667D0 (en) * 2016-01-29 2016-03-16 Ge Healthcare Bio Sciences Ab Improvements in and relating to liquid fraction collectors
WO2017136468A1 (en) * 2016-02-01 2017-08-10 Bio-Rad Laboratories, Inc. Direct contact instrument calibration system
CN105754854B (en) * 2016-03-30 2017-11-24 舟山医院 A kind of stomach cancer cell culture instrument
WO2017197025A1 (en) * 2016-05-11 2017-11-16 Siemens Healthcare Diagnostics Inc. Probe wash station for analytical instrumentation
EP3454987B1 (en) * 2016-05-12 2021-06-30 Siemens Healthcare Diagnostics Inc. Clinical analyzer probe crash detection mechanism and process
WO2018094245A1 (en) * 2016-11-18 2018-05-24 Redbud Labs Small volume sample collection device and related systems and methods
US10427162B2 (en) 2016-12-21 2019-10-01 Quandx Inc. Systems and methods for molecular diagnostics
EP3381560A1 (en) 2017-03-28 2018-10-03 Eppendorf AG Method and a dosing device for contact dosing of liquids
US10816468B2 (en) 2017-05-26 2020-10-27 Gennext Technologies, Inc. Flash photo-oxidation device and higher order structural analysis
US11433402B2 (en) 2017-07-19 2022-09-06 Amgen Inc. Magnetic assisted separation apparatuses and related methods
US10399048B2 (en) * 2017-08-03 2019-09-03 Agilent Technologies, Inc. Sample processing apparatus with integrated heater, shaker and magnet
JP2019066254A (en) * 2017-09-29 2019-04-25 株式会社安川電機 Dispensation system and method for dispensation
CN108181474B (en) * 2017-12-27 2020-07-03 上海合商科学仪器有限公司 Continuous detection analyzer
AU2018353924A1 (en) 2017-12-29 2019-07-18 Clear Labs, Inc. Automated priming and library loading device
EP3520893B1 (en) * 2018-02-02 2020-07-22 F. Hoffmann-La Roche AG System for the thermally controlled processing of a biological sample
EP3549486B1 (en) * 2018-04-06 2020-12-16 Tecan Trading AG Work table for a laboratory automation system and laboratory automation system including said work table
US11506677B2 (en) * 2018-12-21 2022-11-22 Opentrons LabWorks Inc. Systems and methods for pipette robots
EP3971577A4 (en) * 2019-05-13 2023-03-29 Hitachi High-Tech Corporation Automatic analysis apparatus and cleaning method for same
WO2021097100A1 (en) * 2019-11-12 2021-05-20 Amadou Alpha Sall Microsystem label for sample tubes
USD917063S1 (en) 2019-11-20 2021-04-20 Agilent Technologies, Inc. Sample processing apparatus
USD914231S1 (en) 2019-11-20 2021-03-23 Agilent Technologies, Inc. Sample processing apparatus
CN115151652A (en) 2020-02-25 2022-10-04 Dna斯克瑞普特公司 Methods and apparatus for enzymatic synthesis of polynucleotides
US11499244B2 (en) * 2020-02-27 2022-11-15 Marketech International Corp. Device for impregnation using electrophoresis
IL299164A (en) 2020-06-16 2023-02-01 Dna Script Systems, apparatus and kits for enzymatic polynucleotide synthesis
WO2022040598A1 (en) * 2020-08-21 2022-02-24 Beckman Coulter, Inc. Systems and methods for framing workspaces of robotic fluid handling systems
WO2023170258A1 (en) 2022-03-11 2023-09-14 Dna Script Apparatus for enzymatic synthesis of a plurality of polynucleotides comprising a condensation trap
WO2023170286A2 (en) 2022-03-11 2023-09-14 Dna Script Alignment post and secure mechanism for enzymatic polynucleotide synthesis
WO2023170266A1 (en) 2022-03-11 2023-09-14 Dna Script Automation station for enzymatic polynucleotide synthesis
WO2023170259A1 (en) 2022-03-11 2023-09-14 Dna Script Modular accessory rack

Citations (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2579724A (en) * 1946-04-15 1951-12-25 Breakstone Seymour Valved closure plug for insertion in the neck of a bottle
US3754444A (en) * 1970-09-14 1973-08-28 Bio Logics Products Medical sampling and reading
US4224278A (en) * 1977-04-22 1980-09-23 Vitatron Scientific B.V. Apparatus for performing tests and measurements on liquid samples
US4326851A (en) * 1980-10-24 1982-04-27 Coulter Electronics, Inc. Level sensor apparatus and method
WO1983001912A1 (en) * 1981-11-30 1983-06-09 Suovaniemi, Osmo Safety device for sealing a test tube
US4422151A (en) * 1981-06-01 1983-12-20 Gilson Robert E Liquid handling apparatus
US4515752A (en) * 1982-06-18 1985-05-07 Miramanda Fernando X Stopper for containers for use in analyses
US4586546A (en) * 1984-10-23 1986-05-06 Cetus Corporation Liquid handling device and method
EP0209490A2 (en) * 1985-07-08 1987-01-21 ARS Holding 89 N.V. Container for determining the quantity of antibodies or antigens in a biological liquid
US4659677A (en) * 1983-05-26 1987-04-21 Eastman Kodak Company Method providing liquid mixing outside containers
US4730631A (en) * 1985-07-22 1988-03-15 Sequoia-Turner Corporation Probe wash station
US4818492A (en) * 1986-03-20 1989-04-04 Kabushiki Kaisha Toshiba Capacitive liquid level sensor for automatic chemical analyzer
US4820497A (en) * 1986-06-23 1989-04-11 E. I. Du Pont De Nemours And Company Movable cleaning assembly for an aspirating needle
US4869114A (en) * 1987-12-04 1989-09-26 Fuji Photo Film Co., Ltd. Liquid depositing device and method
US4895650A (en) * 1988-02-25 1990-01-23 Gen-Probe Incorporated Magnetic separation rack for diagnostic assays
US4931402A (en) * 1986-03-06 1990-06-05 Tecan Ag Analytische Instrumente Photometric analysis equipment
US4963493A (en) * 1989-10-16 1990-10-16 Daftsios Athanasios C Extraction rack
US4969993A (en) * 1989-08-31 1990-11-13 The Perkin-Elmer Corporation Chromatographic instrument with command sequencing

Family Cites Families (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1180665A (en) * 1915-11-29 1916-04-25 Randall Faichney Company Inc Closure or stopper for serum-containers, &c.
US4134512A (en) * 1977-06-08 1979-01-16 Becton, Dickinson And Company One-way evacuated tube stopper
US4362977A (en) * 1980-06-30 1982-12-07 International Business Machines Corporation Method and apparatus for calibrating a robot to compensate for inaccuracy of the robot
US4322216A (en) * 1981-02-26 1982-03-30 Beckman Instruments, Inc. Method and apparatus for positioning cars in a sample handling apparatus
US4444799A (en) * 1981-08-18 1984-04-24 Nabisco, Inc. Method and composition for producing soft edible baked products and an edible firm gel for use therein
CA1220168A (en) * 1983-09-09 1987-04-07 Henry J. Rahn Magnetic separator for solid phase immunoassays
US4535818A (en) * 1983-09-26 1985-08-20 Vernay Laboratories, Inc. Valve assembly
US4539855A (en) * 1984-05-03 1985-09-10 Eastman Kodak Company Apparatus for transferring liquid out of a capped container, and analyzer utilizing same
CA1268814A (en) * 1984-06-13 1990-05-08 Larry Wayne Moore Apparatus and methods for fluid level sensing
US4566493A (en) * 1985-02-21 1986-01-28 Vernay Laboratories, Inc. Valve assembly
JP2553064B2 (en) * 1987-02-09 1996-11-13 株式会社東芝 Dispensing device
US4906432B1 (en) * 1987-07-17 1991-06-25 Liquid handling
JP2582795B2 (en) * 1987-08-10 1997-02-19 株式会社東芝 Liquid level detector
JP2761385B2 (en) * 1988-04-08 1998-06-04 東亜医用電子株式会社 Immunoagglutination measuring device
EP0342155A3 (en) * 1988-05-13 1990-06-27 Agrogen-Stiftung Laboratory device for optional heating and cooling
US5027075A (en) * 1989-09-18 1991-06-25 Nova Biomedical Corporation Apparatus for determination of probe contact with a liquid surface

Patent Citations (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2579724A (en) * 1946-04-15 1951-12-25 Breakstone Seymour Valved closure plug for insertion in the neck of a bottle
US3754444A (en) * 1970-09-14 1973-08-28 Bio Logics Products Medical sampling and reading
US4224278A (en) * 1977-04-22 1980-09-23 Vitatron Scientific B.V. Apparatus for performing tests and measurements on liquid samples
US4326851A (en) * 1980-10-24 1982-04-27 Coulter Electronics, Inc. Level sensor apparatus and method
US4422151A (en) * 1981-06-01 1983-12-20 Gilson Robert E Liquid handling apparatus
WO1983001912A1 (en) * 1981-11-30 1983-06-09 Suovaniemi, Osmo Safety device for sealing a test tube
US4515752A (en) * 1982-06-18 1985-05-07 Miramanda Fernando X Stopper for containers for use in analyses
US4659677A (en) * 1983-05-26 1987-04-21 Eastman Kodak Company Method providing liquid mixing outside containers
US4586546A (en) * 1984-10-23 1986-05-06 Cetus Corporation Liquid handling device and method
EP0209490A2 (en) * 1985-07-08 1987-01-21 ARS Holding 89 N.V. Container for determining the quantity of antibodies or antigens in a biological liquid
US4730631A (en) * 1985-07-22 1988-03-15 Sequoia-Turner Corporation Probe wash station
US4931402A (en) * 1986-03-06 1990-06-05 Tecan Ag Analytische Instrumente Photometric analysis equipment
US4818492A (en) * 1986-03-20 1989-04-04 Kabushiki Kaisha Toshiba Capacitive liquid level sensor for automatic chemical analyzer
US4820497A (en) * 1986-06-23 1989-04-11 E. I. Du Pont De Nemours And Company Movable cleaning assembly for an aspirating needle
US4869114A (en) * 1987-12-04 1989-09-26 Fuji Photo Film Co., Ltd. Liquid depositing device and method
US4895650A (en) * 1988-02-25 1990-01-23 Gen-Probe Incorporated Magnetic separation rack for diagnostic assays
US4969993A (en) * 1989-08-31 1990-11-13 The Perkin-Elmer Corporation Chromatographic instrument with command sequencing
US4963493A (en) * 1989-10-16 1990-10-16 Daftsios Athanasios C Extraction rack

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Volume 109 No. 25, issued 19 December 1988, WILSON et al., "Automation of Dideoxy-nucleotide DNA Sequencing Reactions Using a Robotic Workstation" see page 225, column 1, Abstract No. 22394d, Biotechniques 6(8) 776-787. *
CHEMICAL AND ENGINEERING NEWS, issued 13 November 1989, STU BORMAN, "New Instrumentation to Speed DNA Sequencing" pg 6. *

Cited By (113)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5410699A (en) * 1989-08-25 1995-04-25 International Business Machines Corp. Apparatus and method for loading BIOS from a diskette in a personal computer system
US5496517A (en) * 1989-12-22 1996-03-05 Beckman Instruments, Inc. Laboratory workstation using thermal vaporization control
EP0505004A2 (en) * 1991-03-21 1992-09-23 Johnson & Johnson Clinical Diagnostics, Inc. Tip to surface spacing for optimum dispensing
EP0505004A3 (en) * 1991-03-21 1992-12-30 Eastman Kodak Company Tip to surface spacing for optimum dispensing
WO1992022800A1 (en) * 1991-06-13 1992-12-23 Abbott Laboratories Liquid dispensing mechanism
EP0555739A1 (en) * 1992-02-13 1993-08-18 F. Hoffmann-La Roche Ag Automatic pipetting device
US5443792A (en) * 1992-02-13 1995-08-22 Hoffmann-La Roche Inc. Pipetting device
WO1993020612A2 (en) * 1992-04-02 1993-10-14 Baxter Deutschland Gmbh Automatic device for the photometric analysis of liquid samples
WO1993020612A3 (en) * 1992-04-02 1993-12-23 Baxter Deutschland Automatic device for the photometric analysis of liquid samples
WO1993025912A2 (en) * 1992-06-09 1993-12-23 Medical Research Council Automated preparation of nucleic acids
WO1993025912A3 (en) * 1992-06-09 1994-03-17 Medical Res Council Automated preparation of nucleic acids
WO1994008759A1 (en) * 1992-10-16 1994-04-28 Thomas Jefferson University Method and apparatus for robotically performing sanger dideoxynucleotide dna sequencing reactions
US5455008A (en) * 1992-10-16 1995-10-03 Thomas Jefferson University Apparatus for robotically performing sanger dideoxynucleotide DNA sequencing reactions using controlled pipet
DE4416406A1 (en) * 1993-05-10 1994-11-17 Olympus Optical Co Output device and output method
EP0707726A1 (en) * 1993-07-09 1996-04-24 Akzo Nobel N.V. Memory control device for an assay apparatus
EP0707726A4 (en) * 1993-07-09 1998-07-15 Akzo Nobel Nv Memory control device for an assay apparatus
US5482864A (en) * 1993-09-17 1996-01-09 Hoffman-La Roche Inc. Method for suspending particles
US5482863A (en) * 1993-09-17 1996-01-09 Hoffmann-La Roche Inc. Apparatus for suspending particles
EP0644426A1 (en) * 1993-09-17 1995-03-22 F. Hoffmann-La Roche Ag Analyser with a device for suspending particles, and suspension method therefor
WO1995011083A2 (en) * 1993-10-22 1995-04-27 Abbott Laboratories Reaction tube and method of use to minimize contamination
WO1995011083A3 (en) * 1993-10-22 1995-08-31 Abbott Lab Reaction tube and method of use to minimize contamination
EP0681160A1 (en) * 1994-05-02 1995-11-08 F. Hoffmann-La Roche AG Analyser with automatic checking of the straightness of a pipetting needle
EP0681184A1 (en) * 1994-05-02 1995-11-08 F. Hoffmann-La Roche AG Analyser with automatic determination of the reference position of a pipetting needle
US5512247A (en) * 1994-05-02 1996-04-30 Hoffmann-La Roche Inc. Apparatus for testing pipetting needle linearity in an automated analyzer
US5529754A (en) * 1994-05-02 1996-06-25 Hoffmann-La Roche Inc. Apparatus for capacitatively determining the position of a pipetting needle within an automated analyzer
WO1997015809A1 (en) * 1995-10-27 1997-05-01 Dynex Technologies (Guernsey) Ltd. Level sensor and washer unit
DE19626234A1 (en) * 1996-06-29 1998-01-02 Innova Gmbh Device for the contamination-free supply and removal of liquids
US6060022A (en) * 1996-07-05 2000-05-09 Beckman Coulter, Inc. Automated sample processing system including automatic centrifuge device
US6058764A (en) * 1997-08-28 2000-05-09 Hitachi, Ltd. Analytical apparatus, liquid chromatography analyzer and a method therefor
JPH1172498A (en) * 1997-08-28 1999-03-16 Hitachi Ltd Analyser
EP0899566A2 (en) * 1997-08-28 1999-03-03 Hitachi, Ltd. Analytical apparatus, liquid chromatography analyzer and a method therefor
EP0899566A3 (en) * 1997-08-28 1999-08-18 Hitachi, Ltd. Analytical apparatus, liquid chromatography analyzer and a method therefor
US8569020B2 (en) 1998-05-01 2013-10-29 Gen-Probe Incorporated Method for simultaneously performing multiple amplification reactions
US8546110B2 (en) 1998-05-01 2013-10-01 Gen-Probe Incorporated Method for detecting the presence of a nucleic acid in a sample
US9598723B2 (en) 1998-05-01 2017-03-21 Gen-Probe Incorporated Automated analyzer for performing a nucleic acid-based assay
US9150908B2 (en) 1998-05-01 2015-10-06 Gen-Probe Incorporated Method for detecting the presence of a nucleic acid in a sample
US8883455B2 (en) 1998-05-01 2014-11-11 Gen-Probe Incorporated Method for detecting the presence of a nucleic acid in a sample
US8569019B2 (en) 1998-05-01 2013-10-29 Gen-Probe Incorporated Method for performing an assay with a nucleic acid present in a specimen
EP0961118A2 (en) * 1998-05-25 1999-12-01 Basf Aktiengesellschaft Pipetting automat
EP0961118A3 (en) * 1998-05-25 2003-05-21 Abbott GmbH & Co. KG Pipetting automat
US6374982B1 (en) 1998-07-14 2002-04-23 Bayer Corporation Robotics for transporting containers and objects within an automated analytical instrument and service tool for servicing robotics
US6332636B1 (en) 1998-07-14 2001-12-25 Bayer Corporation Robotics for transporting containers and objects within an automated analytical instrument and service tool for servicing robotics
US6293750B1 (en) 1998-07-14 2001-09-25 Bayer Corporation Robotics for transporting containers and objects within an automated analytical instrument and service tool for servicing robotics
WO2000056872A3 (en) * 1999-03-22 2000-12-28 Pangene Corporation High-throughput gene cloning and phenotypic screening
WO2000056872A2 (en) * 1999-03-22 2000-09-28 Pangene Corporation High-throughput gene cloning and phenotypic screening
EP1081234A3 (en) * 1999-09-06 2005-04-13 Toyo Boseki Kabushiki Kaisha Apparatus for purifying nucleic acids and proteins
EP1081234A2 (en) * 1999-09-06 2001-03-07 Toyo Boseki Kabushiki Kaisha Apparatus for purifying nucleic acids and proteins
US6986848B2 (en) 1999-09-06 2006-01-17 Toyo Boseki Kabushiki Kaisha Apparatus for purifying nucleic acids and proteins
EP1164200A2 (en) * 2000-05-31 2001-12-19 Pfizer Products Inc. Method and device for drug-drug interaction testing sample preparation
EP1164200A3 (en) * 2000-05-31 2004-02-18 Pfizer Products Inc. Method and device for drug-drug interaction testing sample preparation
WO2002005939A1 (en) * 2000-07-18 2002-01-24 Basf Aktiengesellschaft Method and device for carrying out the automated preparation and characterization of liquid multi-constituent systems
US6827478B2 (en) 2000-07-18 2004-12-07 Basf Aktiengesellschaft Method and device for carrying out the automated preparation and characterization of liquid multi-constituent systems
WO2002027035A3 (en) * 2000-09-28 2003-08-28 Pangene Corporation High-throughput gene cloning and phenotypic screening
WO2002027035A2 (en) * 2000-09-28 2002-04-04 Pangene Corporation High-throughput gene cloning and phenotypic screening
EP1354185A1 (en) * 2001-01-24 2003-10-22 Gilson, Inc. Probe tip alignment for precision liquid handler
EP1354185A4 (en) * 2001-01-24 2010-11-17 Gilson Inc Probe tip alignment for precision liquid handler
DE10241007A1 (en) * 2002-09-05 2004-03-18 Eppendorf Ag Distributing liquid samples from automated liquid stations to automated workstations, for use in molecular biology, comprises that original and target matrix layouts are established to give locations/target destinations
DE10241007B4 (en) * 2002-09-05 2007-08-23 Eppendorf Ag Method for distributing liquid samples
US8021495B2 (en) 2005-04-21 2011-09-20 Wako Pure Chemical Industries, Ltd. Pipette cleaning device and cleaning method
EP1881330A3 (en) * 2006-07-20 2010-10-06 Ortho-Clinical Diagnostics, Inc. Fluid metering in a metering zone
EP1881330A2 (en) * 2006-07-20 2008-01-23 Ortho-Clinical Diagnostics, Inc. Fluid metering in a metering zone
DE102006034245C5 (en) * 2006-07-21 2014-05-28 Stratec Biomedical Systems Ag Positioning device for positioning pipettes
US7976794B2 (en) 2006-07-21 2011-07-12 Stratec Biomedical Systems Ag Positioning device for the positioning of pipettes
JP2010508500A (en) * 2006-10-31 2010-03-18 ビィウルケルト ヴェルケ ゲゼルシャフト ミット ベシュレンクテル ハフツング ウント コンパニー カーゲー Modular laboratory equipment for liquid analysis and synthesis and methods for liquid analysis and synthesis
WO2008052758A1 (en) * 2006-10-31 2008-05-08 Bürkert Werke GmbH & Co. KG Modular laboratory apparatus for analysis and synthesis of liquids and method for analysis and synthesis of liquids
WO2008087138A1 (en) * 2007-01-16 2008-07-24 Roche Diagnostics Gmbh Collection of liquid analytical samples for clinical analytical purpose
EP1947463A1 (en) * 2007-01-16 2008-07-23 Roche Diagnostics GmbH Collection of liquid analytical samples for clinical analytical purpose
US8475740B2 (en) 2007-05-15 2013-07-02 Hitachi High-Technologies Corporation Liquid dispensing apparatus
EP1992952A3 (en) * 2007-05-15 2009-05-06 Hitachi High-Technologies Corporation Liquid dispensing apparatus
EP2565260A2 (en) * 2010-04-30 2013-03-06 Bioneer Corporation Automatic biological sample purification device having a magnetic-field-applying unit, a method for extracting a target substance from a biological sample, and a protein expression and purification method
EP2565260A4 (en) * 2010-04-30 2014-03-12 Bioneer Corp Automatic biological sample purification device having a magnetic-field-applying unit, a method for extracting a target substance from a biological sample, and a protein expression and purification method
US9046455B2 (en) 2010-07-23 2015-06-02 Beckman Coulter, Inc. System and method including multiple processing lanes executing processing protocols
US8840848B2 (en) 2010-07-23 2014-09-23 Beckman Coulter, Inc. System and method including analytical units
US8956570B2 (en) 2010-07-23 2015-02-17 Beckman Coulter, Inc. System and method including analytical units
US8962308B2 (en) 2010-07-23 2015-02-24 Beckman Coulter, Inc. System and method including thermal cycler modules
US9519000B2 (en) 2010-07-23 2016-12-13 Beckman Coulter, Inc. Reagent cartridge
US8996320B2 (en) 2010-07-23 2015-03-31 Beckman Coulter, Inc. System and method including analytical units
US8932541B2 (en) 2010-07-23 2015-01-13 Beckman Coulter, Inc. Pipettor including compliant coupling
US9285382B2 (en) 2010-07-23 2016-03-15 Beckman Coulter, Inc. Reaction vessel
US9140715B2 (en) 2010-07-23 2015-09-22 Beckman Coulter, Inc. System and method for controlling thermal cycler modules
US9274132B2 (en) 2010-07-23 2016-03-01 Beckman Coulter, Inc. Assay cartridge with reaction well
US9529008B2 (en) 2011-03-03 2016-12-27 Life Technologies Corporation Sampling probes, systems, apparatuses, and methods
WO2012119118A3 (en) * 2011-03-03 2016-01-14 Life Technologies Corporation Sampling probes, systems, apparatuses, and methods
US10890596B2 (en) 2011-03-03 2021-01-12 Life Technologies Corporation Sampling probes, systems, apparatuses, and methods
US10082518B2 (en) 2011-03-03 2018-09-25 Life Technologies Corporation Sampling probes, systems, apparatuses, and methods
EP2535712A1 (en) * 2011-06-15 2012-12-19 F. Hoffmann-La Roche AG Analytical system for the preparation of biological material
US10921338B2 (en) 2011-09-09 2021-02-16 Gen-Probe Incorporated Sample container handling with automated cap removal/replacement and drip control
US10132821B2 (en) 2011-09-09 2018-11-20 Gen-Probe Incorporated Automated method for determining the presence of a mucoid strand
US10877057B2 (en) 2011-09-09 2020-12-29 Gen-Probe Incorporated Thermal printing on wall of tubular vessel
US9335336B2 (en) 2011-09-09 2016-05-10 Gen-Probe Incorporated Automated sample handling instrumentation, systems, processes, and methods
US11614454B2 (en) 2011-09-09 2023-03-28 Gen-Probe Incorporated Automated container capping/decapping mechanism
US11815522B2 (en) 2011-09-09 2023-11-14 Gen-Probe Incorporated Automated sample handing instrumentation, systems, processes, and methods
US9506943B2 (en) 2011-11-07 2016-11-29 Beckman Coulter, Inc. Aliquotter system and workflow
US10048284B2 (en) 2011-11-07 2018-08-14 Beckman Coulter, Inc. Sample container cap with centrifugation status indicator device
US9482684B2 (en) 2011-11-07 2016-11-01 Beckman Coulter, Inc. Centrifuge system and workflow
US9910054B2 (en) 2011-11-07 2018-03-06 Beckman Coulter, Inc. System and method for processing samples
US10274505B2 (en) 2011-11-07 2019-04-30 Beckman Coulter, Inc. Robotic arm
US8973736B2 (en) 2011-11-07 2015-03-10 Beckman Coulter, Inc. Magnetic damping for specimen transport system
US9046506B2 (en) 2011-11-07 2015-06-02 Beckman Coulter, Inc. Specimen container detection
US9446418B2 (en) 2011-11-07 2016-09-20 Beckman Coulter, Inc. Robotic arm
CN103111828A (en) * 2013-03-12 2013-05-22 苏州大学 Thin-wall structural member automatic arranging and positioning machine and arranging and positioning method before finish machining
CN103111828B (en) * 2013-03-12 2015-09-30 苏州大学 Thin-walled workpiece auto arrangement localization machine and arrangement localization method before fine finishining
EP3080616B1 (en) 2013-12-12 2020-06-03 Diagnostica Stago Method of determining the position of at least one cartography token
EP3080616B2 (en) 2013-12-12 2023-11-15 Diagnostica Stago Method of determining the position of at least one cartography token
US10201897B2 (en) 2015-09-25 2019-02-12 Kabushiki Kaisha Yaskawa Denki Method for controlling robot and robot system
EP3147669A1 (en) * 2015-09-25 2017-03-29 Kabushiki Kaisha Yaskawa Denki Method for controlling pipetting robot and pipetting robot system
CN108593944A (en) * 2018-03-22 2018-09-28 广州市第人民医院(广州消化疾病中心、广州医科大学附属市人民医院、华南理工大学附属第二医院) Liquid drop chip immunoassay system and method
EP3563929A1 (en) * 2018-04-30 2019-11-06 Firmenich SA Apparatus for customized production of a flavoring agent mix
CN110877771A (en) * 2018-09-06 2020-03-13 里程碑公司 System and method for filling a closed container with a fixative solution
EP3620772A1 (en) * 2018-09-06 2020-03-11 Milestone S.r.l. System and method for filling a closed container with a fixative solution
CN110877771B (en) * 2018-09-06 2021-10-29 里程碑公司 System and method for filling a closed container with a fixative solution
CN110305773A (en) * 2019-08-22 2019-10-08 深圳市芯思微生物科技有限公司 A kind of nucleic acid-extracting apparatus and extracting method
CN112191207A (en) * 2020-09-28 2021-01-08 苏州市启献智能科技有限公司 Chemical experiment system of Internet of things

Also Published As

Publication number Publication date
EP0478753B1 (en) 1997-07-02
DE69126690T2 (en) 1998-01-02
ATE154981T1 (en) 1997-07-15
EP0478753A1 (en) 1992-04-08
US5443791A (en) 1995-08-22
EP0478753A4 (en) 1993-04-07
DE69126690D1 (en) 1997-08-07

Similar Documents

Publication Publication Date Title
EP0478753B1 (en) Automated molecular biology laboratory
EP1506413B1 (en) Automated system for isolating, amplyifying and detecting a target nucleic acid sequence
US8148163B2 (en) Sample processing units, systems, and related methods
EP3662291B1 (en) Sample processing apparatus with integrated heater, shaker and magnet
US9027730B2 (en) Microplate handling systems and related computer program products and methods
US20080026472A1 (en) Instrument For Efficient Treatment Of Analytical Devices
CA2379969A1 (en) Low volume chemical and biochemical reaction system
US8550694B2 (en) Mixing cartridges, mixing stations, and related kits, systems, and methods
Meldrum A biomechatronic fluid-sample-handling system for DNA processing
CN111575172A (en) Gene detection system
EP1615037A1 (en) An apparatus for liquid handling with multiple transfer tools
Mayrand et al. Automation of specific human gene detection

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU NL SE

WWE Wipo information: entry into national phase

Ref document number: 1991908369

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1991908369

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 1991908369

Country of ref document: EP