WO1994021379A1 - Plaque de microtitrage perfectionnee et immunodosages menes a bien dans celle-ci - Google Patents

Plaque de microtitrage perfectionnee et immunodosages menes a bien dans celle-ci Download PDF

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Publication number
WO1994021379A1
WO1994021379A1 PCT/CA1994/000158 CA9400158W WO9421379A1 WO 1994021379 A1 WO1994021379 A1 WO 1994021379A1 CA 9400158 W CA9400158 W CA 9400158W WO 9421379 A1 WO9421379 A1 WO 9421379A1
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WO
WIPO (PCT)
Prior art keywords
reflective
wells
walls
coating
microtitre plate
Prior art date
Application number
PCT/CA1994/000158
Other languages
English (en)
Inventor
Robert Edward Burrell
Original Assignee
Westaim Technologies 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 Westaim Technologies Inc. filed Critical Westaim Technologies Inc.
Priority to AU62561/94A priority Critical patent/AU6256194A/en
Publication of WO1994021379A1 publication Critical patent/WO1994021379A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates

Definitions

  • Microtitre plates are moulded plastic plates or trays having a plurality of depressions or wells in which chemical and biological tests are carried out.
  • Immunogenic assays involving an antibody-antigen reaction to detect an antibody or antigen in biological materials such as serum are most commonly performed in microtitre plates.
  • Colorimetric immunoassays are widely used, the most common type being an ELISA (enzyme-linked immunosorbent assay) method. This technique is an enzyme immunoassay used for the detection of antibodies or antigens.
  • Antigens are immobilized on the surfaces of the wells of the microtitre plates, following which, the sample to be assayed for particular antibodies, typically serum, is added to the wells in specific assay dilutions. This step is followed by the immuno detection of the antibody/antigen complexes, usually by photometrically measuring an enzymatically labelled colour-forming immune complex.
  • a source of incident light is directed onto or through the assay reagents (typically in a clear multi-well microtitre plate) . A portion of the radiant energy of the incident beam is absorbed by a component the assay.
  • the light which is transmitted from the assay is detected by a suitable photometer to provide a measurement of the concentration of a component of the assay.
  • Colorimetric immunoassays are photon absorbing assays.
  • Other photometric assay techniques performed in microtitre plates are termed photon emitting immunoassays. These include chemiluminescent, bioluminescent and fluorescent immunoassays. In these assays photons are emitted, for example as a result of the action of an enzyme, or through the use of particular photon emitting labels. Emitted photons are detected by suitable photometric devices such as luminometers or fluorometers.
  • Immunoassays are typically conducted in microtitre plates.
  • Such plates are typically made of a light transmitting plastic polymer such as polystyrene or polyvinylchoride. It is usually important that the microtitre plate be formed from a light transmitting plastic since reading of the assay results is typically done through the contents in the wells, particularly for colorimetric immunoassays.
  • the microtitre plates may be made of opaque plastic, such as black or white polystyrene, in order to reduce "cross-talk" in photometrically reading the results from well to well (i.e. to reduce interference caused by stray photons). While cross-talk may be lessened by such opaque microtitre plates, the sensitivity of the results is lessened.
  • microtitre plate which provides for improved sensitivity in photometric immunoassay techniques, particularly for photon emitting immunoassays such as, chemiluminescent, bioluminescent and fluorescent immunoassays.
  • microtitre plate with a multiplicity of wells or depressions with walls that are highly reflective.
  • the plates may be formed with a reflective metal, ceramic or semiconductor coating on one or both of the side or bottom walls of the wells.
  • the plates may be formed from reflective metal.
  • Still another alternative is to place a clear microtitre plate inside a reflective plate to read the assay results.
  • Preferred reflective metals, ceramics or semi- conductors for coatings include (in a highly reflective form) aluminum, tin, magnesium, zinc, cadmium, indium, transition metals such as silver, chromium, gold, platinum and nickel, silicon, germanium, silica and alumina or alloys containing at least one of these metals.
  • the plates may also be stamped from a highly reflective metal, for example, from bright aluminum foil.
  • the invention also provides an improved method of conducting a photon emitting immunoassay in the reflective plates. Enhanced sensitivity results from the use of the reflective plates.
  • the invention provides a microtitre plate comprising a plate member having top and bottom surfaces, the top surface defining a plurality of spaced, upwardly opening wells, each of said wells having side and/or bottom walls with inner and outer surface, and one or both of the side and bottom walls of said wells providing a reflective, metallized surface.
  • the invention also provides an improved method of conducting a photon emitting immunoassay in a multi-well microtitre plate, wherein an antigen-antibody complex is formed in the wells and photons emitted from the wells are detected by a photometric device.
  • the improvement comprises conducting the immunoassay in a microtitre plate having a reflective metallized surface on one or both of the side or bottom walls of the wells to reflect photons emitted from the wells into the photometric device.
  • wells as used herein and in the claims, in association with microtitre plates, is meant to include both depressions having only a bottom wall (ex. shallow, round bottom depressions) , and generally cylindrical wells, having both side and bottom walls.
  • Generally cylindrical wells typically have round, flat or conical bottom walls.
  • the side walls are typically vertical, but could vary, for example as conical or fluted side walls.
  • walls as used herein and in the claims is meant to include the inner or outer surfaces of the wells, regardless of the well shape.
  • wall when used in association with a depression shaped well, refers to its rounded surface, although the depression does not in fact possess discrete side and bottom walls.
  • reflective, metallized as used herein and in the claims, is meant to include highly reflective surfaces produced from shiny metallic materials, for instance, aluminum, tin, transition metals, ceramics and semiconductors (all in a highly reflective form) , as distinct from surfaces that are merely light scattering due to glassy or glossy properties.
  • microtitre plates in accordance with the present invention are preferably produced by coating standard multi-well microtitre plates with a reflective, metallized coating on at least a portion of the inner or outer side or bottom walls of the plates.
  • the plates according to this invention are formed by stamping the wells into a reflective metal, such as bright aluminum foil.
  • a standard clear microtitre plate containing the assay inside a second microtitre dish having a reflective metallized coating on the inner side and/or bottom wells.
  • the reflective metallized coatings are preferably metal coatings because they are highly reflective and inexpensive to apply.
  • ceramic or semiconductor materials may be applied as highly reflective coatings in assay applications demanding an inert surface or a surface with particular immobilization qualities.
  • the coating may be provided on the inner side and/or bottom walls of the plates.
  • the outer side and/or bottom walls of the plates are coated.
  • the coating is provided on the inner side and/or bottom walls.
  • the particular areas coated will vary with the particular assay technique and photometric plate reading equipment which is to be used. For instance, for bottom read plates, the bottoms of the wells may be left uncoated.
  • Bottom read plates may include a reflective lid or plate cover for use in the actual reading equipment.
  • Top read plates which are most commonly used, are preferably coated on both the bottom and side walls to maximize the advantages of the reflective coating.
  • the walls of the microtitre plates may be shaped to maximize the utility of the reflective coating.
  • the walls may be curved (ex. rounded or parabolic) or facetted to maximize upward reflections of photons into the photometric detecting equipment.
  • State of the art coating techniques for example physical vapour deposition, chemical vapour deposition or electroless deposition, may be used.
  • Physical vapour deposition techniques include sputtering, magnetron sputtering, vacuum deposition and ion plating. The coating is applied to a thickness such that the walls of the plate are no longer transparent to light, but are highly reflective of light.
  • Microtitre plates which are coated on the outside walls with the metallized reflective coating are preferred to minimize immobilization problems encountered in the assay techniques.
  • An outer reflective coating preserves the inner wall surface (typically polystyrene) for which most immunoassay techniques have been designed. If the inner walls are coated with the metallized reflective coating, the assay technique may be altered to overcome any immobilization problems.
  • a clear, inner coating of a plastic such as polystyrene may be provided on the walls themselves to avoid toxicity or immobilization problems with the metal coating. Reflective ceramic or semiconductor coatings may also be advantageously used to overcome toxicity or immobilization problems.
  • Shiny metals can be used for the reflective metallized coating, for example, aluminum, tin, magnesium, zinc, cadmium, indium, transition metals such as silver, nickel, gold, platinum and chromium or alloys of these or other metals.
  • Aluminum is particularly preferred since it does not interfere significantly with the assay techniques.
  • the particular coating material or alloy chosen will vary with the assay technique to be performed in the wells. Immunoassay techniques are well known in the art.
  • the metallized reflective plates of the present invention are advantageously used in colorimetric immunoassays such as ELISA, to enhance the sensitivity of the readings off the plates.
  • the plates are particulary useful in photon emitting immunoassays, including chemiluminescent, biolu inescent and fluorescent immunoassays.
  • immunoassays including chemiluminescent, biolu inescent and fluorescent immunoassays.
  • assays are described in detail in the literature, see for example, L.J. Kricka and T.J.N. Carter, Clinical and Biochemical Luminescence, Marcel Dekker, New York (1982) ; L.J.Kricka et al. , Analytical Applications of Bioluminescence and Chemiluminescence, Academic Press, New York (1984); and N. Monji and A.
  • the assays may be conducted without alteration. However, if the coating is on the inner walls of the plate, and immobilization or toxicity problems are encountered with the metal coating, the assay technique should be modified to include a first step of coating the inner walls with an adherent plastic film. A reflective coating of ceramic or semiconductor material may also be used.
  • Example 1 This example illustrates the preparation of a metallized microtitre plate in accordance with the present invention.
  • a standard clear flat bottomed polystyrene microtitre plate obtained from Corning was coated by magnetron sputtering with aluminum metal as follows: Magnetron Sputtering Conditions: Equipment - Perkin Elmer 4410 Target - 99.999% Aluminum Power - lkW Time - 20 min R.P.M.
  • Example 2 The two plates coated in accordance with the procedure set forth in Example 1 (i.e. metallized reflective coating on the inside or outside side and bottom walls of the wells) were tested both biologically and physically to demonstrate utility for immunoassays and to evaluate for reflectivity.
  • a) Reflectivity Tests The reflectivity of the each of the plates was compared to the reflectivity of standard clear, white and black microtitre plates using a Nanospec AFT reflectometer. The instrument was standardized against an aluminized optically flat silicon wafer (coated under the conditions set forth in Example 1) . Data was collected as a percentage of the standard.
  • the results were as follows: Standard 100% Plate, metallized inner walls 98% Plate, metallized outer walls 96% Plate, clear polystyrene 9.0% Plate, white polystyrene 7.5% Plate, black polystyrene 4.0%
  • the metallized plates in accordance with the present invention were clearly much more reflective than the clear or opaque, coloured plates. This is in part due to the lack of scattering and absorption of light in the metallized plates relative to the clear and opaque, coloured plates. This demonstrates that in any photon emitting assay, the metallized plates will result in a greater capture of emitted photons since photons not directed at the detector initially will be redirected with minimal loss, even after multiple reflections within the plate.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Optical Measuring Cells (AREA)

Abstract

L'invention concerne une plaque de microtitrage constituée d'une pluralité de puits conçus pour réaliser des immunodosages, une ou plusieurs des parois latérales, de fond et de couvercle de chaque puits présentant une surface métallisée réfléchissante. Ladite face peut être un revêtement interne des parois des plaques transparentes ou opaques, ou un revêtement extérieur des parois de plaques transparentes. Dans un autre mode de réalisation, la plaque peut être formée, par exemple, par estampage à partir d'un matériau métallique réfléchissant. La sensibilité de la détection photométrique est améliorée grâce aux immunodosages à émission de photons réalisés dans la plaque.
PCT/CA1994/000158 1993-03-16 1994-03-11 Plaque de microtitrage perfectionnee et immunodosages menes a bien dans celle-ci WO1994021379A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU62561/94A AU6256194A (en) 1993-03-16 1994-03-11 Enhanced microtitre plate and immunoassays conducted therein

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US3204593A 1993-03-16 1993-03-16
US08/032,045 1993-03-16

Publications (1)

Publication Number Publication Date
WO1994021379A1 true WO1994021379A1 (fr) 1994-09-29

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PCT/CA1994/000158 WO1994021379A1 (fr) 1993-03-16 1994-03-11 Plaque de microtitrage perfectionnee et immunodosages menes a bien dans celle-ci

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AU (1) AU6256194A (fr)
CA (1) CA2158340A1 (fr)
WO (1) WO1994021379A1 (fr)

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0816827A2 (fr) * 1996-06-24 1998-01-07 Schweizerische Eidgenossenschaft vertreten durch das AC-Laboratorium Spiez der Gruppe für Rüstungsdienste Assemblage à cavités multiples pour l'analyse instrumentale
WO1999049974A1 (fr) * 1998-04-01 1999-10-07 The Du Pont Merck Pharmaceutical Company Dispositif sous forme de plaque servant a contenir de petits volumes de liquide
US6027695A (en) * 1998-04-01 2000-02-22 Dupont Pharmaceuticals Company Apparatus for holding small volumes of liquids
US6051191A (en) * 1996-11-25 2000-04-18 Porvair Plc Microplates
EP1024363A2 (fr) * 1999-01-29 2000-08-02 Bodenseewerk Perkin-Elmer Gmbh Détermination quantitative des analytes dans un système hétérogène
DE10060560A1 (de) * 2000-05-26 2001-12-06 Bruker Optik Gmbh Mikrotiterplatte für Infrarotmessungen
WO2003000419A2 (fr) * 2001-06-21 2003-01-03 Hybaid Limited Plaque a puits a echantillons
WO2003025553A2 (fr) * 2001-09-14 2003-03-27 Leybold Optics Gmbh Appareil d'analyse permettant de determiner la structure chimique et/ou la composition d'une pluralite d'echantillons et porte-echantillons
US6800491B2 (en) 2001-06-08 2004-10-05 Nalge Nunc International Corporation Robotic reservoir without liquid hangup
DE10321472A1 (de) * 2003-05-13 2004-12-16 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Fluidik-Modul und Verfahren zu seiner Herstellung
EP1529567A3 (fr) * 2003-11-07 2005-05-25 Herbener, Heinz-Gerd Porte échantillon avec chambre de reaction
WO2006082035A1 (fr) * 2005-02-01 2006-08-10 Greiner Bio-One Gmbh Lame a microresaux a capacite elevee, a base de polymere
EP2098293A1 (fr) 2008-02-22 2009-09-09 Qiagen Instruments AG Plaque de microtitrage
US10317391B2 (en) 2014-06-12 2019-06-11 Axion Biosystems, Inc. Multiwell microelectrode array with optical stimulation

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6266141A (ja) * 1985-09-19 1987-03-25 Sumitomo Bakelite Co Ltd 螢光免疫測定法用容器
WO1989012838A1 (fr) * 1988-06-17 1989-12-28 Wallac Oy Compteur de scintillations liquides de plaquettes-echantillons
WO1991004482A1 (fr) * 1989-09-19 1991-04-04 Park Pharmaceuticals Inc. Luminometre
JPH05203562A (ja) * 1992-01-28 1993-08-10 Hitachi Chem Co Ltd 発光反応用容器及びそれを用いた発光測定法
US5298753A (en) * 1992-11-12 1994-03-29 Wallac Oy Arrangement for counting liquid scintillation samples on bottom-window multi-well sample plates
EP0589691A2 (fr) * 1992-09-23 1994-03-30 Wallac Oy Procéde de prévention de l'infuence optique entre des échantillons de scintillations liquides dans des plaquettes-échantillons translucides

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6266141A (ja) * 1985-09-19 1987-03-25 Sumitomo Bakelite Co Ltd 螢光免疫測定法用容器
WO1989012838A1 (fr) * 1988-06-17 1989-12-28 Wallac Oy Compteur de scintillations liquides de plaquettes-echantillons
WO1991004482A1 (fr) * 1989-09-19 1991-04-04 Park Pharmaceuticals Inc. Luminometre
JPH05203562A (ja) * 1992-01-28 1993-08-10 Hitachi Chem Co Ltd 発光反応用容器及びそれを用いた発光測定法
EP0589691A2 (fr) * 1992-09-23 1994-03-30 Wallac Oy Procéde de prévention de l'infuence optique entre des échantillons de scintillations liquides dans des plaquettes-échantillons translucides
US5298753A (en) * 1992-11-12 1994-03-29 Wallac Oy Arrangement for counting liquid scintillation samples on bottom-window multi-well sample plates

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DATABASE WPI Week 8620, Derwent World Patents Index; AN 86-129702 *
DATABASE WPI Week 8718, Derwent World Patents Index; AN 87-125114 *
DATABASE WPI Week 9336, Derwent World Patents Index; AN 93-285786 *
PATENT ABSTRACTS OF JAPAN vol. 017, no. 626 (P - 1647) 18 November 1993 (1993-11-18) *
PATENT ABSTRACTS OF JAPAN vol. 11, no. 261 (P - 609)<2708> 25 August 1987 (1987-08-25) *

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0816827A3 (fr) * 1996-06-24 1998-06-03 Schweizerische Eidgenossenschaft vertreten durch das AC-Laboratorium Spiez der Gruppe für Rüstungsdienste Assemblage à cavités multiples pour l'analyse instrumentale
EP0816827A2 (fr) * 1996-06-24 1998-01-07 Schweizerische Eidgenossenschaft vertreten durch das AC-Laboratorium Spiez der Gruppe für Rüstungsdienste Assemblage à cavités multiples pour l'analyse instrumentale
US6051191A (en) * 1996-11-25 2000-04-18 Porvair Plc Microplates
WO1999049974A1 (fr) * 1998-04-01 1999-10-07 The Du Pont Merck Pharmaceutical Company Dispositif sous forme de plaque servant a contenir de petits volumes de liquide
US6027695A (en) * 1998-04-01 2000-02-22 Dupont Pharmaceuticals Company Apparatus for holding small volumes of liquids
EP1024363A3 (fr) * 1999-01-29 2004-11-03 Bodenseewerk Perkin-Elmer Gmbh Détermination quantitative des analytes dans un système hétérogène
EP1024363A2 (fr) * 1999-01-29 2000-08-02 Bodenseewerk Perkin-Elmer Gmbh Détermination quantitative des analytes dans un système hétérogène
US7011955B1 (en) 1999-01-29 2006-03-14 Universitaet Tuebingen Quantitative determination of analytes in a heterogeneous system
DE10060560A1 (de) * 2000-05-26 2001-12-06 Bruker Optik Gmbh Mikrotiterplatte für Infrarotmessungen
US6800491B2 (en) 2001-06-08 2004-10-05 Nalge Nunc International Corporation Robotic reservoir without liquid hangup
WO2003000419A2 (fr) * 2001-06-21 2003-01-03 Hybaid Limited Plaque a puits a echantillons
WO2003000419A3 (fr) * 2001-06-21 2003-05-08 Hybaid Ltd Plaque a puits a echantillons
WO2003025553A3 (fr) * 2001-09-14 2003-10-16 Leybold Optics Gmbh Appareil d'analyse permettant de determiner la structure chimique et/ou la composition d'une pluralite d'echantillons et porte-echantillons
WO2003025553A2 (fr) * 2001-09-14 2003-03-27 Leybold Optics Gmbh Appareil d'analyse permettant de determiner la structure chimique et/ou la composition d'une pluralite d'echantillons et porte-echantillons
DE10321472A1 (de) * 2003-05-13 2004-12-16 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Fluidik-Modul und Verfahren zu seiner Herstellung
DE10321472B4 (de) * 2003-05-13 2005-05-12 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Fluidik-Modul, Verfahren zu seiner Herstellung und Verfahren zum Betreiben eines Fluidik-Moduls
EP1529567A3 (fr) * 2003-11-07 2005-05-25 Herbener, Heinz-Gerd Porte échantillon avec chambre de reaction
WO2006082035A1 (fr) * 2005-02-01 2006-08-10 Greiner Bio-One Gmbh Lame a microresaux a capacite elevee, a base de polymere
EP2098293A1 (fr) 2008-02-22 2009-09-09 Qiagen Instruments AG Plaque de microtitrage
US10317391B2 (en) 2014-06-12 2019-06-11 Axion Biosystems, Inc. Multiwell microelectrode array with optical stimulation
US10989703B2 (en) 2014-06-12 2021-04-27 Axion Biosystems, Inc. Multiwell microelectrode array with optical stimulation

Also Published As

Publication number Publication date
CA2158340A1 (fr) 1994-09-29
AU6256194A (en) 1994-10-11

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