WO2003052428A1 - Three-dimensional microfluidics incorporating passive fluid control structures - Google Patents
Three-dimensional microfluidics incorporating passive fluid control structures Download PDFInfo
- Publication number
- WO2003052428A1 WO2003052428A1 PCT/US2002/004045 US0204045W WO03052428A1 WO 2003052428 A1 WO2003052428 A1 WO 2003052428A1 US 0204045 W US0204045 W US 0204045W WO 03052428 A1 WO03052428 A1 WO 03052428A1
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- Y10T137/2267—Device including passages having V over gamma configuration
Definitions
- microfluidic systems are 2 or 2 V. - D, meaning they are made up of microfluidic structures such as channels or wells that lie a single plane. While microfluidic structures in 2 or 2 Vi - D systems have depths, which may vary somewhat from structure to structure, the structures do not vary significantly in elevation with respect to each other, nor does one structure ever cross over or overlap another structure. 2 or 2 V 2 - D systems are prevalent for the simple reason that open channels or wells can be readily formed in a surface of a bulk substrate, and subsequently enclosed by covering the surface of the substrate with a cover plate or film; since substrate surfaces are typically planar, this approach results in the formation of a substantially planar enclosed microfluidic circuit. In contrast, to form structures that overlap or have varying altitudes in a bulk substrate, it is necessary to form at least one of the structures in the interior of the bulk substrate, which is considerably more difficult than forming surface structures.
- Still another object of the invention is to provide a multi-layered microfluidic device capable of mating to conventional substrates such as slides or microtiter plates. This provides the advantage of integrating microfluidic pre- and post-processing capabilities with reactions carried out on or in conventional substrates with microvolumes of fluid.
- FIG. 1 is an exploded view of a multi-layer device incorporating layers including active elements and microfluidic circuitry;
- FIG. 3A depicts a multi-layer device for performing serial dilutions and ELISA
- FIG. 3B is a schematic of the basic microfluidic circuitry of the device of FIG. 3 A;
- FIG. 4 is a multi-layer device for processing a sample and delivering it to a microarray slide with tliree different hybridization solutions
- FIG. 5 is a top view of overlapping channels in a multi-layer microfluidic structure
- FIG. 7 is a top view of overlapping channels in an alternative multi-layer microfluidic structure
- FIG. 8 is a cross-sectional view of the structure of FIG. 7, taken along section line 8-8;
- FIG. 12A is an exploded view of a multi-layer structure containing a well
- FIG. 13 is a perspective view of a fluid chamiel with a passive valve, formed in the surface of a substrate;
- FIG. 14 is an exploded view of a passive valve formed in a multi-layer structure;
- FIG. 15 is a cross-sectional view of the assembled passive valve of FIG. 14;
- Tlie basic three-dimensional structure of the present invention is constructed from multiple thin layers of plastic substrate material sealed together in a leak-free and (optionally) reversible manner.
- Layers may be rigid or flexible, but in general are flat and substantially planar when assembled together.
- Microfluidic structures are formed in the surfaces of individual layers, or through the entire thickness of individual layers, by easily-implemented methods such as molding, micromachining, laser abation, or die cutting. Microfluidic structures thus are primarily formed in planes corresponding to the planes of the substrate layers, although certain structures pass through layers. The exact nature of the layers and the method of sealing vary depending on the particular embodiment of the invention.
- the microfluidic circuitry shown in FIG. 1 is designed for performing polymerase chain reaction (PCR) with a DNA sample to detect sequences of interest, but could be used to implement various biochemical reactions, and particularly those which require that sample be subject to one or more heating steps.
- PCR polymerase chain reaction
- Such reactions include, but are not limited, to various reactions used in DNA processing, for example, PCR, which requires multiple heating steps (thermal cycling), or ligase chain reaction (LCR) or rolling circle amplification (RCA) for DNA amplification, or cycle sequencing, all of which use a single isothermal heating step.
- a PCR cocktail containing DNA sample of interest, but without primers, is pumped (with the use of a syringe pump, for example) into inlet 122 of layer 102.
- Alignment rods 142 are preferably spring-loaded to apply compressive force to hold the layers of device 100 together, and threaded to permit them to be screwed into bottom frame 141.
- Various other methods of assembling the layers with alignment rods may also be devised by one of ordinary skill in the art, and the invention is not limited to any specific method.
- Gasket layers may have the same pattern of openings as one of tlie adjacent layers, and simply perform a sealing function, with the depth of the microfluidic structures primarily defined by the adjacent rigid layer, or, as shown here, the gasket may have a pattern of openings that defines microfluidic structures independent of those defined by openings in the adjacent rigid layer, with the depth of the microfluidic structures in the gasket layer defined by the thickness of the gasket layer.
- Gasket layers may be only a few microns thick, or may be considerably thicker, particularly if the gasket layers define microfluidic structures, rather than simply performing a sealing function.
- Single- or double-sided adhesive sheet materials consisting of plastic sheet materials with adhesive on one or both faces, may be used for some or all layers or the device. Such materials may also be die-cut or laser cut.
- the walls of well cavities 540-545 in layers 532-537, respectively, may be sloped to provide well 530 with a smooth interior surface. Sloped walls can be generated with both molding and laser cutting manufacturing techniques.
- Microfluidic channels, chambers, and valves can be combined to perform various fluid processing tasks.
- One basic task is to divide a fluid stream among multiple channels. This task is facilitated by the use of passive valves.
- the flow of fluid in a network of branching channels can be controlled by providing a set of passive valves at each generation of branches, through which the fluid must pass to reach subsequent generations.
- By making each generation of barriers “stronger” than the previous set fluid is made to fill all branches of the current generation before moving into the next generation of channels. This can be accomplished, for example, by making each successive capillary barrier narrower than the previous set.
- a branching circuit formed in a single substrate layer 600 is depicted in FIG. 21.
- first ELISA circuit 324a includes main channel 330a; conjugate well 333a located on side channel 334a and containing lyophilized enzyme-antibody conjugate formed with an antibody specific to the analyte(s) of interest; substrate well 331a located on side channel 332a and containing lyophilized substrate, with which the conjugate will generate a detectable reaction product; read well 310a on diagnostic surface 303; and waste wells 335a, 336a, 337a, and 338a. All wells and channels are located on layer 302 of the microfluidic device except for read well 310a, which is located on diagnostic surface 303 and connected to circuitry on layer 302 by down via 319a and up via 320a.
Abstract
Description
Claims
Priority Applications (5)
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JP2003553266A JP2005513441A (en) | 2001-02-07 | 2002-02-07 | Three-dimensional microfluidics incorporating passive fluid control structures |
US10/467,585 US20040109793A1 (en) | 2002-02-07 | 2002-02-07 | Three-dimensional microfluidics incorporating passive fluid control structures |
KR10-2003-7010429A KR20030090636A (en) | 2001-02-07 | 2002-02-07 | Three-dimensional microfluidics incorporating passive fluid control structures |
AU2002256996A AU2002256996A1 (en) | 2001-02-07 | 2002-02-07 | Three-dimensional microfluidics incorporating passive fluid control structures |
EP02726572A EP1386169A1 (en) | 2001-02-07 | 2002-02-07 | Three-dimensional microfluidics incorporating passive fluid control structures |
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US60/290,209 | 2001-05-11 | ||
US60/313,703 | 2001-08-20 | ||
US09/967,402 US6601613B2 (en) | 1998-10-13 | 2001-09-28 | Fluid circuit components based upon passive fluid dynamics |
US60/339,851 | 2001-12-12 |
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