Inverted orientation for a microplate

• US 20050244932A1
• Filed: 03/22/2005
• Published: 11/03/2005
• Est. Priority Date: 09/19/2003
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Harding, Ian A.
• Granted Patent: US 7,460,223 B2
• Time in Patent Office: Days
• US Class Current: 435/91.100
• CPC Class Codes: B01L 2200/021 Adjust spacings in an array of wells, pipettes or holders, format transfer between arrays of different size or geometry
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2300/022 Transponder chips
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0851 Bottom walls
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/50851 : specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/50853 : with covers or lids
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 9/523 : for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: B01L 9/56 : Means for indicating position of a recipient or sample in an array
• CPC Class Codes: G01N 2035/00148 : Test cards, e.g. Biomerieux or McDonnel multiwell test cards
• CPC Class Codes: G01N 2035/00356 : Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 2035/0405 : manipulating closing or opening means, e.g. stoppers, screw caps, lids or covers
• CPC Class Codes: G01N 2035/0425 : Stacks, magazines or elevators for plates
• CPC Class Codes: G01N 2035/1037 : Using surface tension, e.g. pins or wires
• CPC Class Codes: G01N 21/274 : Calibration, base line adjustment, drift correction
• CPC Class Codes: G01N 21/553 : and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/648 : using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates

Abstract

An inverted microplate for conducting a thermocycled amplification reaction of polynucleotide. The microplate can comprise a main body having a first and second surfaces and a plurality of wells disposed in the first surface. Each of the plurality of wells can comprise a well opening and a well bottom and be sized to receive an assay. A sealing cover can be operably coupled to the first surface of the main body to seal the well openings of the plurality of wells when the main body is inverted so that the assay is in contact with the sealing covering cover.

Claims

1. A microplate for conducting a thermocycled amplification reaction of polynucleotide, said microplate comprising:

a main body having a first and second surfaces;

a plurality of wells disposed in said first surface, each of said plurality of wells having a well opening and a well bottom and sized to receive an assay; and

a sealing cover operably coupled to said first surface of said main body, said sealing covering cover substantially sealing said well openings of said plurality of wells when said plurality of wells contain said assay and said main body is inverted so that said assay is in contact with said sealing covering cover.

2. The microplate according to claim 1 wherein said plurality of wells includes at least about 96 wells.

3. The microplate according to claim 2 wherein said plurality of wells includes at least about 6144 wells.

4. The microplate according to claim 3 wherein said plurality of wells includes at least about 24,576 wells.

5. The microplate according to claim 3 wherein each of said plurality of wells has a first dimension of about 0.7 mm to about 1.2 mm.

6. The microplate according to claim 3 wherein each of said plurality of wells has a well pitch of about 0.75 mm to about 1.50 mm.

7. The microplate according to claim 1 wherein said main body comprises a thermally conductive plastic.

8. The microplate according to claim 7 wherein said main body comprises a carbon filler.

9. The microplate according to claim 8 wherein said main body is further made of polypropylene.

10. The microplate according to claim 7 wherein said thermally conductive plastic comprises polypropylene, polystyrene, polyethylene, polyethyleneterephthalate, styrene, acrylonitrile, cyclic polyolefin, syndiotactic polystyrene, polycarbonate, liquid crystal polymer, or any combination thereof.

11. The microplate according to claim 1, further comprising:

an alignment feature formed on said main body.

12. The microplate according to claim 11 wherein said alignment feature comprises a convex feature formed on one of said main body and said sealing covering cover, said convex feature operable to engage and align with a concave feature formed on the other of said main body and said sealing covering cover.

13. The microplate according to claim 12 wherein said concave feature is selected from the group consisting essentially of pins, ridges, snaps, and screws.

14. The microplate according to claim 11 wherein said alignment feature comprises a keyed corner on said main body.

15. The microplate according to claim 1, further comprising:

at least one primer and probe disposed in at least one of said plurality of wells.

16. The microplate according to claim 15 wherein said at least one primer and probe are coated to said at least one of said plurality of wells.

17. The microplate according to claim 16, further comprising:

a buffer coated to said at least one of said plurality of wells.

18. The microplate according to claim 1 wherein said main body includes a marking indicia.

19. The microplate according to claim 18 wherein said marking indicia is a barcode.

20. The microplate according to claim 1 wherein said sealing cover includes a pressure sensitive adhesive.

21. The microplate according to claim 20 wherein said pressure sensitive adhesive is a silicone adhesive having low fluorescence.

22. The microplate according to claim 1 wherein said well bottom is substantially hemispherical in shape.

23. The microplate according to claim 1 wherein said plurality of wells is disposed in an array defined by at least about 64 columns and at least 96 rows of wells.

24. The microplate according to claim 1 wherein said first surface being about 127 mm in said first dimension and about 85 mm in said second dimension.

25. The microplate according to claim 1 wherein said cover is substantially rigid.

26. The microplate according to claim 1 wherein said cover is optically transparent.

27. The microplate according to claim 1 wherein said cover comprises a material selected from the group consisting essentially of glass, silicon, quartz, nylon, polystyrene, polyethylene, polypropylene, polytetrafluoroethylene, polycarbonate, polycyclic olefin, cellulose acetate, metal and combinations thereof.

28. The microplate according to claim 1 wherein said cover comprises a thermally conductive material.

29. The microplate according to claim 1, further comprising:

a plurality of primers and probes disposed in at least one of said plurality of wells to perform multiplex PCR.

30. A microplate for conducting a thermocycled amplification reaction of polynucleotide in a liquid sample, said microplate comprising:

a main body having a first and second surfaces;

a plurality of wells disposed in said first surface, each of said plurality of wells having a well opening and a well bottom, each of said plurality of wells being sized to receive the liquid sample; and

a cover having a cover surface, said cover surface being disposed adjacent to said first surface of said main body to cover said top of each of said plurality of wells,

wherein said main body is oriented so that said liquid is in contact with said cover.

31. The microplate according to claim 30, further comprising:

a sealing cover operably disposed between said cover and said first surface.

32. The microplate according to claim 30, further comprising:

at least one primer and probe disposed in at least one of said plurality of wells.

33. The microplate according to claim 30, further comprising:

a plurality of primers and probes disposed in at least one of said plurality of wells to perform multiplex PCR.

34. The microplate according to claim 30, further comprising:

a forward PCR primer, a reverse PCR primer, a FAM labeled MGB quenched, a PCR probe and a buffer disposed in at least one of said plurality of wells.

35. The microplate according to claim 30, further comprising:

a genomic polynucleotides mixture disposed in at least one of said plurality of wells.

36. The microplate according to claim 35 wherein said genomic polynucleotides mixture is from a mammal.

37. The microplate according to claim 36 wherein said mammal is a human.

38. The microplate according to claim 35 wherein said genomic polynucleotides mixture is from a plant.

39. The microplate according to claim 35 wherein said genomic polynucleotides mixture is from a bacterium.

40. The microplate according to claim 35 wherein said genomic polynucleotides mixture is from a species selected from the group consisting essentially of human, mouse, rat, yeast, primate, bacteria, insect, dog, fungus, and virus species.

41. The microplate according to claim 30 wherein said main body comprises a thermally conductive plastic.

42. The microplate according to claim 41 wherein said main body comprises a carbon filler.

43. The microplate according to claim 42 wherein said main body is further made of polypropylene.

44. The microplate according to claim 41 wherein said thermally conductive plastic includes a polymer selected from the group consisting essentially of polypropylene, polystyrene, polyethylene, polyethyleneterephthalate, styrene, acrylonitrile, cyclic polyolefin, syndiotactic polystyrene, polycarbonate, liquid crystal polymer, and mixtures thereof.

45. The microplate according to claim 30 wherein said main body includes a marking indicia.

46. The microplate according to claim 30, further comprising:

an alignment feature formed on said main body.

47. The microplate according to claim 30 wherein said plurality of wells includes at least about 6144 wells.

48. The microplate according to claim 30 wherein said plurality of wells includes at least about 24,576 wells.

49. The microplate according to claim 30 wherein each of said plurality of wells has a first dimension of about 0.7 mm to about 1.2 mm.

50. The microplate according to claim 30 wherein each of said plurality of wells has a well pitch of about 0.75 mm to about 1.50 mm.

51. The microplate according to claim 30 wherein said plurality of wells is disposed in an array defined by at least about 64 columns and at least 96 rows of wells.

52. The microplate according to claim 30 wherein said first surface being about 127 mm in said first dimension and about 85 mm in said second dimension.

53. The microplate according to claim 30 wherein said cover is optically transparent.

54. The microplate according to claim 30 wherein said cover comprises a material selected from the group consisting essentially of glass, silicon, quartz, nylon, polystyrene, polyethylene, polypropylene, polytetrafluoroethylene, polycarbonate, polycyclic olefin, cellulose acetate, metal and combinations thereof.

55. The microplate according to claim 30 wherein said cover comprises a thermally conductive material.

56. A method for performing thermocycling amplification of a liquid polynucleotide sample, said method comprising:

(a) loading a quantity of said liquid polynucleotide sample into a microplate, said microplate having a main body having a first and second surfaces, and a plurality of wells disposed in said first surface, each of said plurality of wells having a well opening and a well bottom, each of said plurality of wells being sized to receive the liquid sample;

(b) covering said main body with a cover, a surface of said cover being disposed adjacent to said first surface of said microplate so as to cover said well openings of said plurality of wells;

(c) inverting said main body such that said liquid polynucleotide sample contacts said cover; and

(d) thermocycling said main body to effect amplification of the polynucleotides in said liquid polynucleotide sample.

57. The method according to claim 56, further comprising:

centrifuging said main body after said inverting said main body.

58. The method according to claim 56 wherein said liquid polynucleotide sample is operable to emit an optical signal during an amplification reaction, said thermocycling said main body further comprises:

(i) placing said inverted main body in contact with a thermal device such that said second surface of said main body is in contact with said thermal device;

(ii) thermocycling said main body to effect said amplification of the polynucleotides in said liquid polynucleotide sample; and

(ii) detecting said optical signal.

59. A kit for performing thermocycling of a liquid polynucleotide sample, said kit comprising:

a microplate having a main body having a first and second surfaces, and a plurality of wells disposed in said first surface, each of said plurality of wells having a well opening and a well bottom; and

a cover having a cover surface, said cover surface connectable to said first surface of said main body to cover said top of each of said plurality of wells;

wherein at least one of said first surface of said main body and said cover surface include a pressure sensitive adhesive.