WO1998046981A1 - Microplateau comportant des puits avec des fonds sureleves - Google Patents

Microplateau comportant des puits avec des fonds sureleves Download PDF

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Publication number
WO1998046981A1
WO1998046981A1 PCT/US1998/007505 US9807505W WO9846981A1 WO 1998046981 A1 WO1998046981 A1 WO 1998046981A1 US 9807505 W US9807505 W US 9807505W WO 9846981 A1 WO9846981 A1 WO 9846981A1
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WO
WIPO (PCT)
Prior art keywords
well
microplate
wells
height
center
Prior art date
Application number
PCT/US1998/007505
Other languages
English (en)
Inventor
Lev J. Leytes
Original Assignee
Ljl Biosystems
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 Ljl Biosystems filed Critical Ljl Biosystems
Priority to EP98918199A priority Critical patent/EP0909381A1/fr
Publication of WO1998046981A1 publication Critical patent/WO1998046981A1/fr

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    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/251—Colorimeters; Construction thereof
    • G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
    • 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
    • G01N21/76—Chemiluminescence; Bioluminescence

Definitions

  • The* present invention relates generally to multi-well bioassay trays , which are commonly referred to as microplates , and in particular to a microplate having wells with elevated bottoms .
  • a microplate is often used to hold mixtures of reagents arid samples to be assayed by techniques which are dependent upon the emission of light signals from the mixtures.
  • the light signals are detected by an optical detector positioned above the microplate.
  • the light signals must be sufficiently strong to be accurately detected. It is also desirable to use a minimum quantity of reagents to reduce the cost of the assay.
  • FIG. 1 shows a cross sectional view of a portion of a conventional microplate 100.
  • Microplate 100 includes a frame 112 having a base 116 and a top portion 120 extending from base 116.
  • Base 116 has a bottom edge 118.
  • Top portion 120 extends to a maximum height HI above bottom edge 118.
  • the current standard for height HI is 14.35 mm.
  • Each well 114 in microplate 100 has sidewalls 122 and a bottom wall 124 attached to sidewalls 122.
  • the top surface of bottom wall 124 forms a well bottom 126.
  • Bottom wall 124 is typically 1.0 mm thick and elevated a height El above bottom edge 118 to prevent scratching of bottom wall 124 when microplate 100 is placed upon a work surface or stacked on top of another microplate.
  • the current standard for height El is 1.0 mm.
  • Well 114 typically has a volume capacity of 300 to 400 microliters for holding a mixture 128 of sample and reagent liquids.
  • a chemical reaction between the sample and reagent liquids causes the generation of a light signal.
  • the light signal travels out of well 114 along a signal path SI to an optical detector 130.
  • Signal path Si has a length LI which is dependent upon the depth of well 114, the depth of mixture 128, and the placement of optical detector 130 above well 114.
  • the depth of well 114 is typically 12.35 mm and optical detector 130 is generally placed 2.0 to 5.0 mm above well 114 to prevent contamination of the detector.
  • the depth of mixture 128 is dependent upon the volume of well 114 occupied by mixture 128.
  • the 384 well plates have the same frame dimensions as the 96 well plates, but the diameter of each well is reduced. The center to center distance between adjacent wells is also reduced, typically to 4.5 mm.
  • the 384 well plates have the same disadvantage of creating a prohibitively long signal path between each sample mixture and an optical detector placed above the mixture.
  • the problem of accurate signal detection is further compounded with 384 well plates because their reduced well diameter creates an even narrower signal path from the bottom of each well to the optical detector. Thus, a significant part of each signal is lost, reducing the sensitivity of the assay.
  • a microplate which reduces the volume of reagent required in each well. It is another object of the invention to provide a microplate which reduces the length of the signal path from the bottom of each well to an optical detector positioned above the microplate, thereby increasing the sensitivity of an assay.
  • the invention presents a microplate for holding mixtures of reagents and samples to be assayed by emission of light signals from the mixtures to an optical detector positioned above the microplate.
  • the microplate includes a frame having a base and a top portion extending from the base.
  • the base has a bottom edge and the top portion extends to a first height above the bottom edge. " The first height is in a range of 12.0 to 16.0 mm, with a preferred value of 14.35 mm so that the microplate is compatible with standard plate handling equipment.
  • the microplate also includes a plurality of wells disposed in the frame for holding the mixtures .
  • the wells are disposed in an eight-by-twelve rectangular array such that the center of each well is spaced 9.0 mm from the center of each horizontally adjacent well and spaced 9.0 mm from the center of each vertically adjacent well. This standard 9.0 mm center to center spacing ensures that the microplate is compatible with conventional liquid transfer equipment.
  • the wells are disposed in an sixteen-by-twenty-four rectangular array such that the center of each well is spaced 4.5 mm from the center of each horizontally adjacent well and spaced 4.5 mm from the center of each vertically adjacent well.
  • Each well has sidewalls and a well bottom.
  • the well bottom is elevated a second height above the bottom edge of the base.
  • the second height is in a range of one half to nine tenths of the first height.
  • the elevation of each well bottom reduces the length of the signal path from each mixture to the optical detector and allows a minimum quantity of reagents to be used in each well.
  • FIG. 1 is a cross sectional view of a portion of a conventional microplate .
  • FIG. 2 is a top plan view of a microplate according to a preferred embodiment of the invention.
  • FIG. 3 is a cross sectional view of the microplate of FIG. 2 taken along the line A - A' in FIG. 2.
  • FIG. 4 is a cross sectional view of a well disposed in the microplate of FIG. 2.
  • FIG. 5 is a cross sectional view of other wells according to the invention.
  • FIG. 6 is a cross sectional view of wells having V-shaped bottoms according to the invention.
  • FIG. 7 is a cross sectional view of wells having U-shaped bottoms according to the invention.
  • FIG. 8 is a cross sectional view of wells having individual filter plug bottoms according to the invention.
  • FIG. 9 is a cross sectional view of wells having filter bottoms formed from a sheet of filter material.
  • FIG. 10 is a top plan view of a microplate having well strips according to the invention.
  • FIG. 11 is a cross sectional view of the microplate of FIG. 10.
  • FIG. 12 is a top plan view of another microplate according to the invention.
  • FIG. 2 shows a top plan view of a microplate 32 having a frame 34 and ninety-six wells 42.
  • Wells 42 are disposed in frame 34 in an eight-by-twelve rectangular array such that the center of each well is spaced a distance C from the center of each horizontally adjacent well.
  • the center of each well is also spaced a distance C from the center of each vertically adjacent well.
  • Distance C is preferably 9.0 mm so that microplate 32 is compatible with standard liquid transfer equipment.
  • FIG. 3 shows a cross section of the microplate taken along the line A - A' in FIG. 2.
  • Frame 34 has a base 36 and a top portion 40 extending from base 36.
  • Base 36 has a bottom edge 38.
  • Top portion 40 extends to a first height H2 above bottom edge 38.
  • Height H2 is preferably in the range of 12.0 to 16.0 mm, with a preferred value of 14.35 mm. The height of 14.35 mm is presently preferred so that the microplate is compatible with standard plate handling equipment.
  • Each well 42 has sidewalls 44 extending vertically to a second height above bottom edge 38.
  • the second height is preferably equal to height H2.
  • Each well also has a bottom wall 46 whose upper surface forms a well bottom 48.
  • each well bottom 48 is flat.
  • Bottom wall 46 is attached to sidewalls 44 such that each well bottom is elevated a third height E2 above bottom edge 38.
  • Height E2 is one half to nine tenths of height H2.
  • height E2 is in the range of 6.0 to 14.4 mm with a preferred range of 7.2 to 12.9 mm when height H2 is equal to 14.35 mm.
  • FIG. 4 shows a detailed cross sectional view of the structure of each well.
  • Each well is designed to hold a mixture 28 of reagent and sample liquids to be assayed by the emission of light signals from mixture 28 to an optical detector 30.
  • Mixture 28 has a depth I which is dependent upon a volume V of mixture 28.
  • Detector 30 is positioned a height G above the well. Height G is selected such that detector 30 is located as close as possible to the well without becoming contaminated by samples and reagents . Height G is typically 2.0 to 5.0 mm with a preferred value of 3.0 mm.
  • each well has a diameter D in the range of 6.0 to 8.0 mm with a preferred value of 7.0 mm. Also in the preferred embodiment, each well has a depth F m the range of 1.5 to 7.1 mm with a preferred value of 3.0 mm.
  • the diameter and depth of each well is preferably selected such that the well has a volume capacity of 100 to 150 microliters.
  • Microplate 32 is easily fabricated using conventional injection molding techniques and is preferably a single molded plastic part.
  • FIG. 4 A chemical reaction between the sample and reagent liquids causes the generation of a light signal.
  • the light signal travels out of the well along a signal path S2 to optical detector 30.
  • volume V ranges from 10 to 45 microliters.
  • length L2 of signal path S2 ranges from 4.80 to 5.75 mm as volume V ranges from 10 to 45 microliters.
  • the light signal generated by a 10 to 45 microliter mixture of sample and reagents is capable of traveling this path length with sufficient strength to be accurately detected by detector 30.
  • microplate 32 provides a significant improvement over conventional microplates by allowing sensitive assays to be performed with a minimum quantity of reagents .
  • the microplate improves the accuracy of the assay while simultaneously reducing costs .
  • a further advantage of microplate 32 is that it achieves both of these goals while maintaining the frame dimensions and well center spacing necessary to be compatible with standard plate handling and liquid transfer equipment.
  • FIG. 5 shows a second embodiment of the invention in which the microplate includes a plurality of wells 50 in place of wells 42.
  • Wells 50 are preferably disposed in frame 34 in the same manner as wells 42, e.g. 96 wells arranged in an 8x12 array on 9.0 mm centers.
  • wells 50 differ from wells 42 in that wells 50 have a smaller volume capacity tor smaller volume applications .
  • Each well 50 has a diameter J in the range of 2.0 to 5.0 mm and a depth K in the range of 1.5 to 4.0 mm.
  • the diameter and depth of each well 50 is preferably selected such that each well has a volume capacity of 10 to 50 microliters.
  • each well bottom 48 is elevated to a height E3 above bottom edge 38.
  • Height E3 is preferably seven tenths to nine tenths of height H2, e.g. 10.0 to 12.9 mm when height H2 is equal to 14.35 mm. This elevation of each well bottom 48 ensures that optical detector 30 receives an adequate signal from mixture 28.
  • the operation and advantages of the second embodiment are the same as the preferred embodiment described above.
  • FIGS. 6 - 7 illustrate alternative embodiments of the microplate in which the well bottoms have different shapes.
  • FIG. 6 shows wells 52 having V-shaped well bottoms.
  • Wells 52 are preferably disposed in frame 34 in . " the same manner as the wells of the preferred embodiment, e.g. 96 wells arranged in an 8x12 array on 9.0 mm centers.
  • wells 52 differ from the wells of the preferred embodiment in that wells 52 have bottom walls 54 whose upper surfaces form V-shaped well bottoms 56. Bottom walls 54 are positioned such that the lowest point of each well bottom 56 is elevated height E2 above edge 38.
  • FIG. 7 shows wells 58 having U-shaped well bottoms.
  • Wells 58 are preferably disposed in frame 34 in the same manner as the wells of the preferred embodiment, e.g. 96 wells arranged in an 8x12 array on 9.0 mm centers.
  • wells 58 differ from the wells of the preferred embodiment in that wells 58 have bottom walls 60 whose upper surfaces form U-shaped well bottoms 62. Bottom walls 60 are positioned such that the lowest point of each well bottom 62 is elevated height E2 above bottom edge 38.
  • FIGS. 8 - 9 illustrate alternative embodiments of the microplate in which the well bottoms are formed by filter material.
  • FIG. 8 shows wells 76 having well bottoms 80 formed by individual filter plugs 78.
  • Wells 76 are preferably disposed in frame 34 in the same manner as the wells of the preferred embodiment, e.g. 96 wells arranged in an 8x12 array on 9.0 mm centers .
  • Filter plugs 78 are positioned such that each well bottom 80 is elevated height E2 above bottom edge 38.
  • Each filter plug is preferably press fit between the sidewalls of a respective well.
  • each filter plug may be ultrasonically bonded or adhesively bonded to the sidewalls of a respective well .
  • One advantage of the current invention is that filter plugs 78 may have a thickness ranging up to height E2 and still fit within frame 34. Filter bottoms are typically thicker than non-filter bottoms, and their greater thickness often makes them difficult to fit to conventional microplates .
  • FIG. 9 shows wells 64 having well bottoms 68 formed by a single filter sheet 70.
  • Wells 64 are preferably disposed in frame 34 in the same manner as the wells of the preferred embodiment, e.g. 96 wells arranged in an 8x12 array on 9.0 mm centers.
  • Sheet 70 is positioned such that each well bottom 68 is elevated height E2 above bottom edge 38.
  • Each well 64 has respective sidewalls 66 to which sheet 70 is attached.
  • Sheet 70 includes barrier sections 82 disposed below the sidewalls of each well. Barrier sections 82 prevent liquid transfer between adjacent wells.
  • Sheet 70 is preferably attached to the sidewalls of each well by ultrasonic bonding. In this process, the top surface of sheet 70 is held firmly pressed to the bottom surfaces of the sidewalls. Sheet 70 is then bonded to the sidewalls by the application of ultrasonic energy which causes the contacting surfaces to fuse together. The fusing of the contacting surfaces forms barrier sections 82.
  • sheet 70 may be press fit and adhesively bonded to frame 34. As sheet 70 is press fit, force of contact between the top surface of sheet 70 and the bottom surfaces of the sidewalls compresses sheet 70 below the contacting surfaces . The compression of sheet 70 forms barrier sections 82.
  • FIGS. 10 - 11 illustrate a third embodiment of the invention.
  • FIG. 10 shows a top plan view of a microplate 84 having a frame 86 and a number of well strips 88 removably disposed in frame 86.
  • Frame 86 has a rim 87 for holding well strips 88.
  • the number of well strips disposed in frame 86 at any given time may be varied to tailor microplate 84 to a specific application.
  • Frame 86 is preferably sized to hold at least eight well strips concurrently.
  • Each well strip 88 forms a single row of wells 90.
  • Wells 90 are preferably formed in each well strip such that the center of each well is spaced 9.0 mm from the center of each adjacent well.
  • Frame 86 and well strips 88 may be easily fabricated using conventional injection molding techniques.
  • FIG. 11 shows a cross sectional view of one of well strips 88 disposed in frame 86.
  • Frame 86 includes a base 36 having a bottom edge 38. Frame 86 extends to height H2 above bottom edge 38.
  • Each well 90 formed in the well strip has sidewalls 92 and a bottom wall 94 whose upper surface forms a well bottom 96.
  • Each well bottom is elevated height E2 above bottom edge 38, as previously described in the preferred embodiment above.
  • the advantage of the third embodiment is that it allows wells to be easily added and removed from the microplate. Other than the differences described, the operation and advantages of the third embodiment are the same as those described in the preferred embodiment above.
  • FIG. 12 shows a top plan view of an alternative microplate 74 having 384 wells disposed in a sixteen-by-twenty-four (16x24) rectangular array.
  • the wells are disposed such that the center of each well is spaced a distance Y from the center of each horizontally and vertically adjacent well. Distance Y is preferably 4.5 mm.
  • Each well in microplate 74 has an elevated well bottom which may be a flat bottom, V-shaped bottom, U-shaped bottom, or filter bottom as previously described.
  • Microplate 74 is easily fabricated using conventional injection molding techniques. The advantage of microplate 74 is that it allows a greater number of samples to be concurrently assayed for high throughput screening applications .
  • the wells have tapered sidewalls to further reduce the volume of reagent required in each well .
  • the wells are square rather than round. Round wells are illustrated in the preferred embodiment due to their predominant use in industry, but the microplate of the present invention is also effective with wells having different shapes. Further, the above embodiments describe wells arranged in 8x12 and 16x24 rectangular arrays. However, the microplate of the present invention may include any number of wells arranged in any format, e.g. 6 wells arranged in a 2x3 array, 24 wells arranged in a 4x6 array, 48 wells arranged in a 6x8 array, etc.

Abstract

Cette invention se rapporte à un microplateau servant à contenir des mélanges (28) de réactifs et des échantillons à doser par l'émission de signaux lumineux à partir des mélanges en direction d'un détecteur optique (30) placé au-dessus du microplateau. Ce microplateau comprend un cadre (34) ayant une base (36) et une partie supérieure (40) s'étendant à partir de la base. Ladite base comporte un bord inférieur (38) et la partie supérieure (40) s'étend jusqu'à une hauteur (H2) au-dessus du bord inférieur (38). Cette hauteur (H2) est de préférence égale à 14,35 mm, pour que ledit microplateau soit compatible avec les équipements de manipulation de plateaux standards. Ce microplateau comprend également plusieurs puits (50) disposés dans le cadre (34) et servant à contenir les mélanges (28). Chaque puits présente des parois latérales (44) et un fond (48). Chaque fond de puits est surélevé à une hauteur (H3) au-dessus du bord inférieur (38) de la base (36). Cette hauteur (H3) est comprise entre la moitié et les neuf dixièmes de la hauteur (H2). La position surélevée de chaque fond de puits permet de réduire la longueur du trajet (S3) des signaux allant de chaque mélange (28) au détecteur optique (30) et permet d'utiliser une quantité minimum de réactifs dans chaque puit.
PCT/US1998/007505 1997-04-14 1998-04-14 Microplateau comportant des puits avec des fonds sureleves WO1998046981A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP98918199A EP0909381A1 (fr) 1997-04-14 1998-04-14 Microplateau comportant des puits avec des fonds sureleves

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US84055397A 1997-04-14 1997-04-14
US08/840,553 1997-04-14

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6254833B1 (en) 1998-02-24 2001-07-03 Aurora Biosciences Corporation Microplate lid
US6721053B1 (en) 1999-05-19 2004-04-13 Corning Intellisense Corporation System for high resolution chemical and biological sensing
US6825042B1 (en) 1998-02-24 2004-11-30 Vertex Pharmaceuticals (San Diego) Llc Microplate lid
WO2005052644A2 (fr) * 2003-11-21 2005-06-09 Perkinelmer Las, Inc. Dispositif optique integre a une cupule
WO2007057468A1 (fr) * 2005-11-21 2007-05-24 Boehringer Ingelheim International Gmbh Procede et systeme complet destines au developpement de formes et a leurs tests in vitro presentant une bonne previsibilite de l'absorption in vivo, un rendement eleve et de faibles exigences en ce qui concerne la substance active
US7271885B2 (en) 2004-03-25 2007-09-18 Perkinelmer Las, Inc. Plasmon resonance measuring method and apparatus
EP2012125A1 (fr) 2000-04-28 2009-01-07 MDS Analytical Technologies (US) Inc. Analyses de modification moléculaire
WO2014121179A1 (fr) * 2013-02-01 2014-08-07 Bio-Rad Laboratories, Inc. Système destiné à l'aspiration d'une émulsion
WO2015200803A1 (fr) * 2014-06-26 2015-12-30 The Trustees Of The University Of Pennsylvania Puces multipuits micro-fabriquées pour formation d'image automatisée à long terme de croissance et de comportement de c elegans

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4704255A (en) * 1983-07-15 1987-11-03 Pandex Laboratories, Inc. Assay cartridge

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4704255A (en) * 1983-07-15 1987-11-03 Pandex Laboratories, Inc. Assay cartridge

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6254833B1 (en) 1998-02-24 2001-07-03 Aurora Biosciences Corporation Microplate lid
US6825042B1 (en) 1998-02-24 2004-11-30 Vertex Pharmaceuticals (San Diego) Llc Microplate lid
US6721053B1 (en) 1999-05-19 2004-04-13 Corning Intellisense Corporation System for high resolution chemical and biological sensing
EP2012125A1 (fr) 2000-04-28 2009-01-07 MDS Analytical Technologies (US) Inc. Analyses de modification moléculaire
US7233391B2 (en) 2003-11-21 2007-06-19 Perkinelmer Las, Inc. Optical device integrated with well
WO2005052644A3 (fr) * 2003-11-21 2006-04-20 Perkinelmer Las Inc Dispositif optique integre a une cupule
WO2005052644A2 (fr) * 2003-11-21 2005-06-09 Perkinelmer Las, Inc. Dispositif optique integre a une cupule
US7271885B2 (en) 2004-03-25 2007-09-18 Perkinelmer Las, Inc. Plasmon resonance measuring method and apparatus
WO2007057468A1 (fr) * 2005-11-21 2007-05-24 Boehringer Ingelheim International Gmbh Procede et systeme complet destines au developpement de formes et a leurs tests in vitro presentant une bonne previsibilite de l'absorption in vivo, un rendement eleve et de faibles exigences en ce qui concerne la substance active
WO2014121179A1 (fr) * 2013-02-01 2014-08-07 Bio-Rad Laboratories, Inc. Système destiné à l'aspiration d'une émulsion
US9388945B2 (en) 2013-02-01 2016-07-12 Bio-Rad Laboratories, Inc. System for emulsion aspiration
WO2015200803A1 (fr) * 2014-06-26 2015-12-30 The Trustees Of The University Of Pennsylvania Puces multipuits micro-fabriquées pour formation d'image automatisée à long terme de croissance et de comportement de c elegans
US11795423B2 (en) 2014-06-26 2023-10-24 The Trustees Of The University Of Pennsylvania Micro-fabricated multi-well chips for long term automated imaging of C. elegans growth and behavior

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