Multi-through hole testing plate for high throughput screening

• US 6,436,632 B2
• Filed: 10/04/2001
• Issued: 08/20/2002
• Est. Priority Date: 03/19/1999
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Liu, Amy Deming, Schellenberger, Volker
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/970,578
• Publication Number:
  US 20020015994A1
• Time in Patent Office: 320 Days
• Field of Search: 435/4, 435/283.1, 435/29, 435/30, 422/50, 422/68.1
• US Class Current: 435/4
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00319 the blocks being mounted in stacked arrangements
• CPC Class Codes: B01J 2219/0052 in the shape of elongated tubes
• CPC Class Codes: B01J 2219/00524 in the shape of fiber bundles
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00662 Two-dimensional arrays within two-dimensional arrays
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/50857 using arrays or bundles of open capillaries for holding samples
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices

Abstract

A method for holding samples for analysis and an apparatus thereof includes a testing plate with a pair of opposing surfaces and a plurality of holes. Each of the holes extends from one of the opposing surfaces to the other one of the opposing surfaces. The holes are arranged in groups, where each group has at least two rows and two columns of holes. The groups are arranged in sets, where each set has at least two rows and two columns of groups. To analyze samples, at least one of the opposing surfaces of the testing plate is immersed in a solution to be analyzed. A portion of the solution enters openings for each of the holes in the immersed opposing surface. Once the holes are filled with solution, the testing plate is removed and is held above a supporting surface. Surface tension holds the solution in each of the holes. The solution in one or more of the holes is then analyzed and the solution in one of these holes is identified for further study. The location of the identified solution is marked based upon its location within a particular set and group of holes.

Claims

1. An assembly for holding liquid samples, said assembly comprising:

a substrate having first and second opposing surfaces and including a bundle of capillary tubes, wherein said first opposing surface comprises first ends of the capillary tubes of said bundle and said second opposing surface comprises said second ends of the capillary tubes of said bundle; and

a plurality of through holes in said substrate, including longitudinally-extending holes through the centers of the capillary tubes of said bundle, each of said through holes extending from an opening in one of the opposing surfaces of the substrate to an opening in the other of the opposing surfaces, wherein the through holes are sized to provide sufficient surface tension by capillary action to hold respective liquid samples.

2. The assembly of claim 1, wherein said bundle further includes a band that bounds the capillary tubes together.

3. The assembly of claim 2, wherein said band comprises metal, plastic, glass, or an elastomeric compound.

4. The assembly of claim 1, wherein each capillary tube of said bundle has a length between the first end and second end thereof that is at least two times greater than the average diameter of said capillary tube.

5. The assembly of claim 1, wherein each capillary tube of said bundle has a length between the first end and second end thereof that is at least four times greater than the average diameter of said capillary tube.

6. The assembly of claim 1, wherein the average diameter of each capillary tube is from about 0.001 millimeter to about one millimeter.

7. The assembly of claim 1, wherein the length of each capillary tube is from about one millimeter to about one centimeter.

8. The assembly of claim 1, wherein each capillary tube has a capacity to hold from about 0.0001 microliter to about ten microliters of liquid sample.

9. The assembly of claim 1, wherein said capillary tubes are fused, bonded or adhered together in said bundle.

10. The assembly of claim 1, wherein said bundle of capillary tubes comprises from about 100 to greater than 1,000 capillary tubes.

11. The assembly of claim 1, wherein said bundle of capillary tubes comprises from about 500 to about 1,500 capillary tubes.

12. The assembly of claim 1, wherein said bundle of capillary tubes has a circular cross-section.

13. The assembly of claim 1, wherein said bundle of capillary tubes comprises a rectangular or square array of capillary tubes.

14. The assembly of claim 1, wherein said bundle of capillary tubes includes capillary tubes arranged in rows and rows of capillary tubes arranged in columns.

15. A method of providing a plurality of liquid samples, said method comprising:

providing the assembly of claim 1,; and

at least partially filling a plurality of said through holes with respective liquid samples, wherein surface tension holds the respective liquid samples in the respective plurality of through holes.

16. The method of claim 15, wherein said at least partially filling comprises contacting at least one of the opposing surfaces of the assembly with at least one liquid sample such that portions of said at least one liquid sample at least partially fill a respective plurality of said through holes.

17. The method of claim 16, further comprising, after said contacting, separating the opposing surfaces of the assembly from said liquid samples such that the surfaces are substantially free of said liquid sample and the portions of said sample held within the respective plurality of through holes are isolated from one another.

18. The method of claim 17, wherein said contacting comprises immersing at least one of said opposing surfaces into said at least one liquid sample.

19. A method for identifying the location of a liquid sample in a testing assembly, said method comprising:

providing the assembly of claim 1 wherein the capillary tubes are arranged in groups, and each of the groups comprises at least two rows of through holes and at least two columns of through holes, and wherein portions of the liquid sample are located in at least some of the through holes;

identifying the liquid sample in at least one through hole; and

locating the identified through hole based upon the group in which the through hole is located.

20. The assembly of claim 1, wherein a plurality of said capillary tubes are filled with respective liquid samples each containing a mixture of biological cells, and wherein at least one of said liquid samples contains at least one analyte cell.

21. A high throughput screening method comprising:

providing at least one liquid sample that includes an analyte that directly or indirectly produces a detectable characteristic;

providing the testing assembly of claim 1;

at least partially filling a plurality of said through holes with at least portions of said at least one liquid sample, wherein surface tension holds the respective portions in the respective plurality of through holes; and

detecting which of said plurality of through holes contains a liquid sample portion that includes said analyte.

22. The method of claim 21, wherein said analyte is a biological cell.

23. The method of claim 21, wherein said detectable characteristic is a fluorescence or absorption characteristic.

24. The method of claim 21, wherein said analyte is at least one of a mutant cell, a secretable protein, an enzyme, or a microorganism.

57. A high throughput screening method comprising:

providing at least one liquid sample that includes an analyte that directly or indirectly produces a detectable characteristic in one or more of an absorbance transcription assay, a fluorescent transcription assay, a fluorescent secreted enzyme assay, and a microorganism screening assay;

providing the assembly of claim 1;

at least partially filling a plurality of said through holes of said assembly with at least portions of said at least one liquid sample, wherein surface tension holds the respective portions in the respective plurality of through holes; and

detecting which of said plurality of through holes contains a liquid sample portion that includes said analyte.

58. The method of claim 57, wherein said detectable characteristic is produced in an absorbance transcription assay.

59. The method of claim 57, wherein said detectable characteristic is produced in a fluorescent transcription assay.

60. The method of claim 57, wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.

61. The method of claim 57, wherein said detectable characteristic is produced in a microorganism screening assay.

25. An assembly for holding liquid samples for analysis, said assembly comprising;

a substrate having a pair of opposing surfaces, and side edges;

a plurality of through holes in the substrate each of said through holes extending from an opening in one of the opposing surfaces of the substrate to an opening in the other of the opposing surfaces, said through holes being sized to provide sufficient surface tension by capillary action to hold respective liquid samples; and

a handle connected to said substrate for maneuvering said substrate.

26. The assembly of claim 25, wherein said handle is connected to said substrate with bolts.

27. The assembly of claim 25, wherein an opening is provided in one of said side edges, and an end of said handle is removably positioned within said opening.

28. The assembly of claim 25, wherein said handle extends out from the side of the substrate.

62. A high throughput screening method comprising:

providing at least one liquid sample that includes an analyte that directly or indirectly produces a detectable characteristic in one or more of an absorbance transcription assay, a fluorescent transcription assay, a fluorescent secreted enzyme assay, and a microorganism screening assay;

providing the assembly of claim 25;

at least partially filling a plurality of said through holes with at least portions of said at least one liquid sample, wherein surface tension holds the respective portions in the respective plurality of through holes; and

detecting which of said plurality of through holes contains a liquid sample portion that includes said analyte.

63. The method of claim 62, wherein said detectable characteristic is produced in an absorbance transcription assay.

64. The method of claim 62, wherein said detectable characteristic is produced in a fluorescent transcription assay.

65. The method of claim 62, wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.

66. The method of claim 62, wherein said detectable characteristic is produced in a microorganism screening assay.

29. An assembly for holding liquid samples for analysis, said assembly comprising:

a substrate having a pair of opposing surfaces;

a plurality of through holes in the substrate, each of said through holes extending from an opening in one of the opposing surfaces of the substrate to an opening in the other of the opposing surfaces, said through holes being sized to provide sufficient surface tension by capillary action to hold respective liquid samples; and

a first evaporation plate secured to one of the opposing surfaces of the substrate, and a second evaporation plate secured to the other of said opposing surfaces of the substrate, said evaporation plates being secured to said substrate to preserve samples dispersed in the through holes from evaporation and contamination.

30. The assembly of claim 29, wherein said evaporation plates are secured to said substrate with clamps.

31. The assembly of claim 29, wherein said evaporation plates are secured to said substrate with bolts.

32. The assembly of claim 29, wherein each of said opposing surfaces includes a recessed portion and said plurality of through holes are located in the recessed portions of the opposing surfaces, and wherein the plurality of through holes in the recessed portions are spaced away from facing surfaces of the evaporation plates when the evaporation plates are secured to the substrate.

67. A high throughput screening method comprising:

providing at least one liquid sample that includes an analyte that directly or indirectly produces a detectable characteristic in one or more of an absorbance transcription assay, a fluorescent transcription assay, a fluorescent secreted enzyme assay, and a microorganism screening assay;

providing the assembly of claim 29;

at least partially filling a plurality of said through holes with at least portions of said at least one liquid sample, wherein surface tension holds the respective portions in the respective plurality of through holes; and

detecting which of said plurality of through holes contains a liquid sample portion that includes said analyte.

68. The method of claim 67, wherein said detectable characteristic is produced in an absorbance transcription assay.

69. The method of claim 67, wherein said detectable characteristic is produced in a fluorescent transcription assay.

70. The method of claim 67, wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.

71. The method of claim 67, wherein said detectable characteristic is produced in a microorganism screening assay.

33. An assembly for holding liquid samples for analysis, said assembly comprising:

a substrate having a pair of opposing surfaces; and

a plurality of through holes in the substrate, each of said through holes extending from an opening in one of the opposing surfaces of the substrate to an opening in the other one of the opposing surfaces of the substrate, said through holes being sized to provide sufficient surface tension by capillary action to hold respective liquid samples,

wherein a recessed portion having a first depth is provided in a first opposing surface of said pair and a protruding portion having a height is provided on the other opposing surface of said pair, and wherein each of said plurality of through holes extends from the recessed portion of said first opposing surface to the protruding portion of the other opposing surface, and said height is less than said depth.

34. A stackable system comprising two or more of the assemblies of claim 33 stacked together, wherein the protruding portion of one of the assemblies is received within the recessed portion of another of the assemblies.

35. A system comprising two of the assemblies of claim 33 stacked together and further comprising a spacer between the two stacked assemblies.

72. A high throughput screening method comprising:

providing at least one liquid sample that includes an analyte that directly or indirectly produces a detectable characteristic in one or more of an absorbance transcription assay, a fluorescent transcription assay, a fluorescent secreted enzyme assay, and a microorganism screening assay;

providing the assembly of claim 33;

at least partially filling a plurality of said through holes with at least portions of said at least one liquid sample, wherein surface tension holds the respective portions in the respective plurality of through holes; and

detecting which of said plurality of through holes contains a liquid sample portion that includes said analyte.

73. The method of claim 72, wherein said detectable characteristic is produced in an absorbance transcription assay.

74. The method of claim 72, wherein said detectable characteristic is produced in a fluorescent transcription assay.

75. The method of claim 72, wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.

76. The method of claim 72, wherein said detectable characteristic is produced in a microorganism screening assay.

36. A system comprising at least a first and a second assembly stacked one on top of the other, each assembly comprising:

a substrate having a first opposing surface and a second opposing surface; and

a plurality of through holes in the substrate, each of said through holes extending from an opening in the first opposing surface of the substrate to an opening in the second opposing surface of the substrate, said through holes being sized to provide sufficient surface tension by capillary action to hold respective liquid sample, and

wherein said system further comprises an evaporation plate disposed between said first assembly and said second assembly, said evaporation plate having a first surface facing the second opposing surface of said first assembly, and said evaporation plate having a second surface facing the first opposing surface of said second assembly, and wherein at least one of the surfaces of said evaporation plate is provided with a recessed portion to space the surface of the recessed portion from ends of the through holes at one or both of the second opposing surface of the first assembly and the first opposing surface of the second assembly.

37. The system of claim 36, wherein the second opposing surface of said first assembly is provided with a recessed portion.

38. The system of claim 36, wherein the first opposing surface of said second assembly is provided with a recessed portion.

39. The system of claim 36, wherein both the first and second surfaces of said evaporation plate are provided with a recessed portion.

40. A system comprising at least a first and a second assembly stacked one on top of the other, each assembly comprising:

a substrate having a first opposing surface and a second opposing surface; and

a plurality of through holes in the substrate, each of said through holes extending from an opening in the first opposing surface of the substrate to an opening in the second opposing surface of the substrate, said through holes being sized to provide sufficient surface tension by capillary action to hold respective liquid sample, and

wherein said system further comprises an evaporation plate disposed between said first assembly and said second assembly, said evaporation plate having a first surface facing the second opposing surface of said first assembly, and said evaporation plate having a second surface facing the first opposing surface of said second assembly, and wherein at least one of the second opposing surface of said first assembly and the first opposing surface of said second is provided with a recessed portion to space ends of the through holes at one or both of the second opposing surface of the first assembly and the first opposing surface of the second assembly, from the evaporation plate.

41. The system of claim 40, wherein the second opposing surface of said first assembly is provided with a recessed portion.

42. The system of claim 40, wherein the first opposing surface of said second assembly is provided with a recessed portion.

43. The system of claim 40, wherein both the second opposing surface of said first assembly and the first opposing surface of said second assembly are provided with a recessed portion.

44. The system of claim 40, wherein at least one of the first and second surfaces of said evaporation plate is provided with a recessed portion.

45. The system of claim 40, wherein a spacer is provide between the evaporation plate and at least one of said first and second assemblies.

46. A system comprising two or more assemblies stacked one on top of the other, each assembly comprising:

a substrate having a first opposing surface and a second opposing surface; and

a plurality of through holes in the substrate, each of said through holes extending from an opening in the first opposing surface of the substrate to an opening in the second opposing surface of the substrate, said through holes being sized to provide sufficient surface tension by capillary action to hold respective liquid sample, and

wherein said system further comprises an evaporation plate disposed between said first assembly and said second assembly, said evaporation plate having a first surface facing the second opposing surface of said first assembly, said evaporation plate having a second surface facing the first opposing surface of said second assembly, and wherein a spacer is provided between at least one of the second surfaces of said evaporation plate and at least one of the first and second assemblies, said spacer spacing openings of the through holes away from the evaporation plate.

47. The system of claim 46, wherein a spacer is disposed between the second opposing surface of said first assembly and the evaporation plate.

48. The system of claim 46, wherein a spacer is disposed between the first opposing surface of said second assembly and the evaporation plate.

49. The system of claim 46, wherein at least one of said surfaces of said evaporation plate is provided with a recessed portion.

50. A plate for holding samples of a solution with biological material for analysis, said plate comprising;

a platform having a pair of opposing surfaces; and

a plurality of holes in the platform, each of said holes extending from an opening in one of the opposing surfaces of the platform to an opening in the other one of the opposing surfaces of the platform, each of the holes being sized to hold biological material in said through holes by surface tension; and

wherein at least one of the opposing surfaces is provided with a recessed portion.

51. The plate of claim 50, wherein both opposing surfaces are provided with a recessed portion.

52. The plate of claim 50, wherein the plurality of holes contains portions of a sample of a biological material, the biological material includes cells, and each of the holes is sized to hold a plurality of the cells.

53. The plate of claim 50, wherein the holes are arranged in groups on the substrate, each of the groups comprises at least two rows of holes and at least two columns of holes.

54. An assembly comprising two or more of the plates of claim 50 stacked one on top of the other.

55. The assembly of claim 54, wherein a spacer is provided between at least two of the plates of said assembly.

56. The assembly of claim 54, wherein at least one of said plates of said assembly is provided with a handle.

77. A high throughput screening method comprising:

providing at least one liquid sample that includes an analyte that directly or indirectly produces a detectable characteristic in one or more of an absorbance transcription assay, a fluorescent transcription assay, a fluorescent secreted enzyme assay, and a microorganism screening assay;

providing the plate of claim 50;

at least partially filling a plurality of said through holes with at least portions of said at least one liquid sample, wherein surface tension holds the respective portions in the respective plurality of through holes; and

detecting which of said plurality of through holes contains a liquid sample portion that includes said analyte.

78. The method of claim 77, wherein said detectable characteristic is produced in an absorbance transcription assay.

79. The method of claim 77, wherein said detectable characteristic is produced in a fluorescent transcription assay.

80. The method of claim 77, wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.

81. The method of claim 77, wherein said detectable characteristic is produced in a microorganism screening assay.