WO1991015751A1 - Method of improving assay sensitivity - Google Patents
Method of improving assay sensitivity Download PDFInfo
- Publication number
- WO1991015751A1 WO1991015751A1 PCT/GB1991/000567 GB9100567W WO9115751A1 WO 1991015751 A1 WO1991015751 A1 WO 1991015751A1 GB 9100567 W GB9100567 W GB 9100567W WO 9115751 A1 WO9115751 A1 WO 9115751A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- optical
- index matching
- fcfd
- waveguide
- edge
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N2021/0346—Capillary cells; Microcells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
- G01N2021/6465—Angular discrimination
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N2021/7769—Measurement method of reaction-produced change in sensor
- G01N2021/7786—Fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/648—Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
-
- Y—GENERAL 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
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/804—Radioisotope, e.g. radioimmunoassay
Definitions
- the present invention relates to a method of improving the sensitivity of optical assays wherein the light signal to be analyzed emerges from the edge of an optical waveguide.
- the invention is particularly applicable to assays carried out using fluorescence capillary fill devices (FCFD's) in which an optical waveguide forms one plate of the device.
- FCFD's fluorescence capillary fill devices
- FCFD's and methods for their manufacture are described in detail in EP-A-171148, whilst photometric methods including assays carried out using FCFD's are described in EP-A-170376.
- Assays involving FCFD's rely on angular separation, in light emerging from the optical waveguide, between fluorescence originating from molecules in sample liquid within the FCFD cavity and fluorescence orginating from molecules bound, directly or indirectly, to the waveguide.
- This angular separation occurs because fluorescent material in solution can only fluoresce into the waveguide at relatively large angles relative to the plane of the waveguide since the angle of any incident beam relative to this plane will be increased by refraction at the solution/waveguide interface.
- Such light consequently emerges from the waveguide at large angles (e.g. > 47° relative to the axis of the waveguide) .
- Fluorescent material bound to the surface of the waveguide emits light into the waveguide at all angles, which light thereafter emerges from the waveguide over a wide range of angles relative to the axis of the waveguide.
- Fluorescent material bound to the surface of the waveguide emits light into the waveguide at all angles, which light thereafter emerges from the waveguide over a wide range of angles relative to the axis of the waveguide.
- FCFD's As described in EP-A-171148, a convenient method of manufacturing FCFD's involves preparation of coated glass 1 sandwiches' from which individual FCFD's are separated by scribing and breaking. This manufacturing process permits the inexpensive bulk production of FCFD's, this being particularly desirable in view of the disposable nature of these devices.
- the optical edge it is difficult within the context of such a bulk production process, however, to design scribing and breaking procedures which consistently produce FCFD's in which the end of the waveguide from which emerging light is detected (hereinafter referred to as "the optical edge") is optically smooth. It will be appreciated that surface irregularities at the optical edge will give rise to some degree of light scattering and consequent mixing of the narrow angle light emission attributable only to surface-bound fluorescent material and the broader angle emission attributable to both surface-bound fluorescent material and fluorescent material in solution. This inevitably degrades the signal quality and overall performance of optical assay techniques using FCFD's.
- the present invention is based on the discovery that light scattering at the optical edge can be substantially reduced in a particularly simple and efficient manner if the optical edge is contacted with an index matching substance so as to avoid having an optical edge/air interface.
- index matching substance is meant a substance having a refractive index similar to that of the material of the optical waveguide, e.g. having a refractive index which is ⁇ 10% that of the waveguide. Since high index glass as normally used in the production of waveguides typically has a refractive index of about 1.5, index matching substances having a refractive index in the range 1.35-1.65 are particularly appropriate for such applications.
- Intimate contact between the optical edge and the index matching substance may be achieved by, for example, selecting an index matching substance which is a liquid or gel, or by employing precursors for a substantially transparent solid which pliably moulds to the surface of the optical edge before subsequently setting or otherwise solidifying.
- Suitable liquid index matching substances include those traditionally employed as immersion fluids in microscopy, such as cedar oil and Canada balsam.
- Other liquids with appropriate refractive indices include silicones, ethyl alcohol, amyl alcohol, aniline, benzene, glycerol, paraffin oil and turpentine.
- Appropriate gels include, for example, silicone gels.
- Suitable precursors for solids include adhesives which may, for example, be selected from appropriate epoxy and acrylate systems, and optical cements as well as plastics materials (including thermoplastics) with appropriate refractive index, for example silane elastomers.
- readily meltable solids of appropriate refractive index e.g. naphthalene, may be applied in molten form and then allowed to cool and solidify.
- the method of the invention may, for example, be effected by index matching the optical edge to a defined optically smooth component, such as an optical flat, which forms the first stage of an optical detector train.
- Index matching may, for example, be made to an optical structure as described in published application WO 90/15985, the contents of which are herein incorporated by reference.
- one or more FCFD's are positioned in a disposable holder (advantageously of the radially extending type described in published patent application WO 90/11830.
- Such holders may if desired incorporate an optical component such as a lens to which the optical edge is matched. Holders so constructed and incorporating an index matching substance constitute a further feature of the invention.
- Fig. 1 is a schematic representation of a portion of a disposable holder incorporating an FCFD
- Fig.2 is a vertical section through part of an alternative form of holder incorporating an FCFD.
- Fig.3 is an enlarged partial view of the cross section of Fig.2.
- upper glass plate 1 and optical wave guide 2 make up an FCFD mounted by means not shown in a disposable holder (e.g. produced by injection moulding), a part of which is shown as 3.
- the design of the holder is such that a plurality of FCFD cells radiate horizontally from a central vertical axis.
- That part of the edge of the holder closest to optical wave guide 2 is configured as a lens 4 associating with optical detector 5.
- Index matching substance 6, which is advantageously a solid such as an optical cement or appropriate plastic which during application pliably moulds to the optical edge 7 of waveguide 2, ensures that scattering of light between optical edge 7 and lens 4 is minimised.
- Example 1 In the configuration shown in Fig. 2 the upper glass plate 1 and optical waveguide 2 which constitute an FCFD are mounted in a holder comprising a base unit 8 and lid 9. A glass window 10 is mounted between said base unit 8 and lid 9 and is matched to the optical edge of wave guide 2 by means of index matching substance 6.
- Example 1 For a better understanding of the invention the following non-limitative Example is provided by way of illustration: Example 1
- the configuration illustrated in Fig.2 is employed.
- the glass window 10 is a flame polished microscope slide (e.g. as manufactured by Chance Propper Ltd., Smethwick, Warley, U.K.) which is cut to appropriate dimensions.
- the height of the glass window is chosen so that those light rays 11, 12 which would normally (i.e. in the absence of index matching material) emerge from the waveguide at angles ⁇ 47° to the axis thereof are refracted by the glass window without being reflected off its edge surface 13.
- a suitable window is 4 mm by 10 mm with a thickness of 1.0 mm.
- the FCFD and glass window are placed in approximately moulded features in the (usually plastic) base unit 8 so that there is a small gap of approximately 0.1 mm between the optical edge 7 of the FCFD and the glass window.
- a drop of index matching material such as a UV-curable optical adhesive (e.g. Norland Optical Adhesive 81, Norland Products Inc. , New Brunswick, U.S.A.) is then applied so that the small gap fills by capillary action and the adhesive is retained between the two components by surface tension.
- the adhesive is then cured by exposure to ultraviolet light at a wavelength of between 320 mm and 400 mm.
- the holder lid 9 is then attached to the base 8 and the whole may be stored for future use.
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP50699791A JP3286314B2 (en) | 1990-04-11 | 1991-04-10 | How to improve assay sensitivity |
US07/930,690 US5369717A (en) | 1990-04-11 | 1991-04-10 | Optical waveguide assay unit and method of improving assay sensitivity using same |
DE69118178T DE69118178T2 (en) | 1990-04-11 | 1991-04-10 | METHOD FOR IMPROVING THE SENSITIVITY OF ANALYZES |
EP91907433A EP0524224B1 (en) | 1990-04-11 | 1991-04-10 | Method of improving assay sensitivity |
CA002078568A CA2078568C (en) | 1990-04-11 | 1991-04-10 | Method of improving assay sensitivity |
GR960400870T GR3019483T3 (en) | 1990-04-11 | 1996-04-02 | Method of improving assay sensitivity |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB909008261A GB9008261D0 (en) | 1990-04-11 | 1990-04-11 | Method of improving assay sensitivity |
GB9008261.1 | 1990-04-11 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1991015751A1 true WO1991015751A1 (en) | 1991-10-17 |
Family
ID=10674305
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/GB1991/000567 WO1991015751A1 (en) | 1990-04-11 | 1991-04-10 | Method of improving assay sensitivity |
Country Status (12)
Country | Link |
---|---|
US (1) | US5369717A (en) |
EP (1) | EP0524224B1 (en) |
JP (1) | JP3286314B2 (en) |
AT (1) | ATE135818T1 (en) |
AU (1) | AU642736B2 (en) |
CA (1) | CA2078568C (en) |
DE (1) | DE69118178T2 (en) |
DK (1) | DK0524224T3 (en) |
ES (1) | ES2084813T3 (en) |
GB (1) | GB9008261D0 (en) |
GR (1) | GR3019483T3 (en) |
WO (1) | WO1991015751A1 (en) |
Cited By (4)
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---|---|---|---|---|
WO1993018405A1 (en) * | 1991-01-30 | 1993-09-16 | Du Pont | Apparatus for performing assays using evanescent waves |
US5677196A (en) * | 1993-05-18 | 1997-10-14 | University Of Utah Research Foundation | Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays |
US5919712A (en) * | 1993-05-18 | 1999-07-06 | University Of Utah Research Foundation | Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays |
US6611634B2 (en) | 1996-03-19 | 2003-08-26 | University Of Utah Research Foundation | Lens and associatable flow cell |
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US5669997A (en) * | 1995-07-13 | 1997-09-23 | Hughes Danbury Optical Systems, Inc. | Method of bonding optical members together |
US5568253A (en) * | 1996-03-20 | 1996-10-22 | Taiwan Semiconductor Manufacturing Company Ltd. | Sample holder for sample testing apparatus |
US6180288B1 (en) | 1997-03-21 | 2001-01-30 | Kimberly-Clark Worldwide, Inc. | Gel sensors and method of use thereof |
US6060256A (en) * | 1997-12-16 | 2000-05-09 | Kimberly-Clark Worldwide, Inc. | Optical diffraction biosensor |
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US7640083B2 (en) | 2002-11-22 | 2009-12-29 | Monroe David A | Record and playback system for aircraft |
US6221579B1 (en) | 1998-12-11 | 2001-04-24 | Kimberly-Clark Worldwide, Inc. | Patterned binding of functionalized microspheres for optical diffraction-based biosensors |
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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 |
US6399295B1 (en) | 1999-12-17 | 2002-06-04 | Kimberly-Clark Worldwide, Inc. | Use of wicking agent to eliminate wash steps for optical diffraction-based biosensors |
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US7223368B2 (en) | 2002-05-03 | 2007-05-29 | Kimberly-Clark Worldwide, Inc. | Diffraction-based diagnostic devices |
US7091049B2 (en) | 2002-06-26 | 2006-08-15 | Kimberly-Clark Worldwide, Inc. | Enhanced diffraction-based biosensor devices |
US7285424B2 (en) | 2002-08-27 | 2007-10-23 | Kimberly-Clark Worldwide, Inc. | Membrane-based assay devices |
US7169550B2 (en) * | 2002-09-26 | 2007-01-30 | Kimberly-Clark Worldwide, Inc. | Diffraction-based diagnostic devices |
US7247500B2 (en) | 2002-12-19 | 2007-07-24 | Kimberly-Clark Worldwide, Inc. | Reduction of the hook effect in membrane-based assay devices |
US8675276B2 (en) * | 2003-02-21 | 2014-03-18 | Kla-Tencor Corporation | Catadioptric imaging system for broad band microscopy |
US7851209B2 (en) | 2003-04-03 | 2010-12-14 | Kimberly-Clark Worldwide, Inc. | Reduction of the hook effect in assay devices |
US20040197819A1 (en) | 2003-04-03 | 2004-10-07 | Kimberly-Clark Worldwide, Inc. | Assay devices that utilize hollow particles |
US7713748B2 (en) | 2003-11-21 | 2010-05-11 | Kimberly-Clark Worldwide, Inc. | Method of reducing the sensitivity of assay devices |
US7943395B2 (en) | 2003-11-21 | 2011-05-17 | Kimberly-Clark Worldwide, Inc. | Extension of the dynamic detection range of assay devices |
US20050112703A1 (en) | 2003-11-21 | 2005-05-26 | Kimberly-Clark Worldwide, Inc. | Membrane-based lateral flow assay devices that utilize phosphorescent detection |
US7943089B2 (en) | 2003-12-19 | 2011-05-17 | Kimberly-Clark Worldwide, Inc. | Laminated assay devices |
US20070196863A1 (en) * | 2006-02-17 | 2007-08-23 | Hanson Technologies, Inc. | Prion protein detection |
US20080026373A1 (en) * | 2006-07-26 | 2008-01-31 | Rodionova Natalia A | Assays Based On Light Emission From Analyte Complexes Within A Cassette |
US9357925B2 (en) * | 2013-08-02 | 2016-06-07 | John Adamovics | Method and apparatus for scanning 3D dosimeters |
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US4124364A (en) * | 1976-12-02 | 1978-11-07 | International Standard Electric Corporation | Method for making glass sleeve fiber splice |
US4469500A (en) * | 1981-01-26 | 1984-09-04 | Rca Corporation | Method of cleaving a crystal to produce a high optical quality corner |
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WO1990001157A1 (en) * | 1988-07-22 | 1990-02-08 | Ord Corp. | Immunoassay apparatus |
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WO1990015985A1 (en) * | 1989-06-22 | 1990-12-27 | Ares-Serono Research & Development Limited Partnership | Method of optical analysis |
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GB8911462D0 (en) * | 1989-05-18 | 1989-07-05 | Ares Serono Res & Dev Ltd | Devices for use in chemical test procedures |
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US5192510A (en) * | 1991-01-30 | 1993-03-09 | E. I. Du Pont De Nemours And Company | Apparatus for performing fluorescent assays which separates bulk and evanescent fluorescence |
-
1990
- 1990-04-11 GB GB909008261A patent/GB9008261D0/en active Pending
-
1991
- 1991-04-10 JP JP50699791A patent/JP3286314B2/en not_active Expired - Fee Related
- 1991-04-10 WO PCT/GB1991/000567 patent/WO1991015751A1/en active IP Right Grant
- 1991-04-10 ES ES91907433T patent/ES2084813T3/en not_active Expired - Lifetime
- 1991-04-10 US US07/930,690 patent/US5369717A/en not_active Expired - Lifetime
- 1991-04-10 AT AT91907433T patent/ATE135818T1/en not_active IP Right Cessation
- 1991-04-10 EP EP91907433A patent/EP0524224B1/en not_active Expired - Lifetime
- 1991-04-10 CA CA002078568A patent/CA2078568C/en not_active Expired - Fee Related
- 1991-04-10 AU AU76689/91A patent/AU642736B2/en not_active Ceased
- 1991-04-10 DK DK91907433.6T patent/DK0524224T3/en active
- 1991-04-10 DE DE69118178T patent/DE69118178T2/en not_active Expired - Fee Related
-
1996
- 1996-04-02 GR GR960400870T patent/GR3019483T3/en unknown
Patent Citations (7)
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US4124364A (en) * | 1976-12-02 | 1978-11-07 | International Standard Electric Corporation | Method for making glass sleeve fiber splice |
US4469500A (en) * | 1981-01-26 | 1984-09-04 | Rca Corporation | Method of cleaving a crystal to produce a high optical quality corner |
WO1987006956A1 (en) * | 1986-05-05 | 1987-11-19 | Battelle Memorial Institute | Analytical method for detecting and measuring specifically sequenced nucleic acids |
WO1988001376A1 (en) * | 1986-08-14 | 1988-02-25 | Radiometer A/S | Method and apparatus for determining the level of an analyte in a sample of whole blood |
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Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993018405A1 (en) * | 1991-01-30 | 1993-09-16 | Du Pont | Apparatus for performing assays using evanescent waves |
US5300423A (en) * | 1991-01-30 | 1994-04-05 | E. I. Du Pont De Nemours And Company | Specific binding assay involving separation of light emissions |
US5677196A (en) * | 1993-05-18 | 1997-10-14 | University Of Utah Research Foundation | Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays |
US5919712A (en) * | 1993-05-18 | 1999-07-06 | University Of Utah Research Foundation | Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays |
US6316274B1 (en) | 1993-05-18 | 2001-11-13 | University Of Utah Research Foundation | Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays |
US6979567B2 (en) | 1993-05-18 | 2005-12-27 | Biocentrex, Llc | Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays |
US6611634B2 (en) | 1996-03-19 | 2003-08-26 | University Of Utah Research Foundation | Lens and associatable flow cell |
Also Published As
Publication number | Publication date |
---|---|
EP0524224B1 (en) | 1996-03-20 |
AU642736B2 (en) | 1993-10-28 |
GR3019483T3 (en) | 1996-07-31 |
DK0524224T3 (en) | 1996-04-15 |
DE69118178T2 (en) | 1996-09-12 |
ES2084813T3 (en) | 1996-05-16 |
DE69118178D1 (en) | 1996-04-25 |
GB9008261D0 (en) | 1990-06-13 |
ATE135818T1 (en) | 1996-04-15 |
US5369717A (en) | 1994-11-29 |
EP0524224A1 (en) | 1993-01-27 |
JP3286314B2 (en) | 2002-05-27 |
AU7668991A (en) | 1991-10-30 |
CA2078568C (en) | 2002-06-11 |
JPH05505874A (en) | 1993-08-26 |
CA2078568A1 (en) | 1991-10-12 |
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