Method and devices for partitioning biological sample liquids into microvolumes

• US 6,391,578 B2
• Filed: 04/02/2001
• Issued: 05/21/2002
• Est. Priority Date: 04/09/1997
• Status: Expired due to Term

• Current Assignee: 3M Innovative Properties Company (3M Company)
• Original Assignee: 3M Innovative Properties Company (3M Company)
• Inventors: Qiu, Jean, Wickert, Peter D., Wei, Ai-Ping, Debe, Mark K., Williams, Michael G., Halverson, Kurt J., Krejcarek, Gary E., Berg, James G., Calhoun, Clyde D.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US09/824,160
• Publication Number:
  US 20010024805A1
• Time in Patent Office: 414 Days
• Field of Search: 435/29, 435/31, 435/34, 435/39, 435/288.4, 435/288.3
• US Class Current: 435/39
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/20 Material Coatings immunocoatings C12M25/00
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 25/04 in combination with well or multiwell plates, i.e. culture inserts
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404

Abstract

A method for partitioning an aqueous biological liquid sample into discrete microvolumes for detection and enumeration of microorganisms is described. The method involves distributing microvolumes of a sample to a plurality of hydrophilic liquid-retaining zones of a culture device, where each liquid-retaining zone is surrounded by a portion of a hydrophobic “land” area. Also disclosed are devices for carrying out these methods.

Claims

1. A culture device for detection or enumeration of microorganisms, said device comprising a substrate comprising liquid-retaining discs comprising a hydrophilic material, wherein said substrate comprises a material that is hydrophobic relative to said liquid-retaining discs and wherein the discs comprise media for growth of microorganisms.

2. The culture device of claim 1 wherein said discs are constructed at least in part of a material selected from the group consisting of cellulosics, polyolefins, polyamides and polyesters.

3. The culture device of claim 2 wherein the discs are constructed at least in part of alpha cellulose.

4. The culture device of claim 2 wherein the discs are constructed at least in part of rayon.

5. The culture device of claim 2 wherein the discs are constructed at least in part nylon.

6. The culture device of claim 2 wherein the discs are constructed at least in part of polylactic acid.

7. The culture device of claim 1 wherein said device has about 10 to about 10,000 discs.

8. The culture device of claim 1, wherein said device has about 400 to about 600 discs.

9. The culture device of claim 1, wherein each said disc has a liquid retention capacity of about 1 to about 2 microliters.

10. The culture device of claim 1, having an indicator substance on said discs.

11. The culture device of claim 1 wherein the discs are adhered to the substrate with adhesive, and wherein said adhesive has an indicator for detecting microorganisms.

12. The culture device of claim 10, wherein said indicator substance is selected from the group consisting of a chromogenic indicator, a fluorescent indicator, a luminescent indicator and an electrochemical indicator.

13. The culture device of claim 1, wherein said culture device comprises a plurality of at least two sets of liquid-retaining discs, each set comprising discs of uniform size, said sets varying in liquid retention capacity.

21. The culture device of claim 1 wherein at least one of the discs has a microvolume capacity of liquid retention.

14. A method for partitioning an aqueous liquid sample into discrete microvolumes, comprising:

a) providing a device for culturing a microorganism, said device comprising an assay surface, said surface comprising liquid-retaining discs comprising a hydrophilic material and a land area comprising a hydrophobic material between said discs, each said disc comprising media for growing microorganisms; and

b) contacting said liquid sample with said assay surface such that said liquid sample is partitioned into said liquid-retaining discs.

15. The method of claim 14, wherein the discs are constructed of a material selected from the group consisting of cellulosics, polyolefins, polyamides and polyesters.

16. The method of claim 15, wherein the discs are coated with a growth medium.

17. The method of claim 14, wherein said discs have at least one indicator substance coated thereon.

18. The method of claim 17, wherein said indicator substance is selected from the group consisting of a chromogenic indicator, a fluorescent indicator, a luminescent indicator and an electrochemical indicator.

19. The method of claim 14, wherein said discs are of uniform size and each said disc has a liquid retention capacity of about 0.01 to about 25 microliters.

20. The method of claim 14 wherein each said disc has a liquid retention capacity of about 1 to about 2 microliters.

22. The method of claim 14 wherein at least one of the discs has a microvolume capacity of liquid retention.

23. A method for partitioning an aqueous liquid sample into discrete microvolumes, comprising:

a) providing a device for culturing a microorganism, the device comprising an assay surface, the assay surface comprising liquid-retaining zones comprising a hydrophilic material and a land area comprising a hydrophobic material between the zones; and

b) contacting the liquid sample with the assay surface such that the liquid sample is partitioned into the liquid-retaining zones.

24. The method of claim 23 wherein at least one of the liquid-retaining zones has a microvolume capacity of liquid retention.

25. The method of claim 23 wherein the zones comprise a coating or deposition of assay reagent.

26. The method of claim 25 wherein the assay reagent comprises nutrient medium.

27. The method of claim 25 wherein the assay reagent comprises at least one indicator substance.

28. The method of claim 27 wherein the indicator substance is a chromogenic indicator, a fluorescent indicator, a luminescent indicator or an electrochemical indicator.

29. The method of claim 23 wherein the zones are uniform size and each zone has a liquid retention capacity of about 0.01 to about 25 microliters.

30. The method of claim 29 wherein each zone has a liquid retention capacity of about 1 to about 2 microliters.

31. The method of claim 23 wherein the culture device comprises about 10 to about 10,000 liquid-retaining zones.

32. The method of claim 31 wherein the culture device comprises about 400 to about 600 liquid-retaining zones.

33. The method of claim 23 wherein the liquid-retaining zones comprise microvolume wells.

34. The method of claim 23 wherein the land area comprises a treated nanostructured film.

35. The method of claim 23 wherein the liquid-retaining zones comprise hydrophilic fiber material projecting from the hydrophobic material.

36. The method of claim 35 wherein at least one of the zones comprises a hydrophilic absorbent disc.

37. The method of claim 35 wherein at least one of the zones comprises protruding hydrophilic nonwoven fiber loop material.

38. The method of claim 23 wherein the culture device comprises a plurality of at least two sets of liquid-retaining zones, each set having zones of uniform size, the sets varying in liquid retention capacity.

39. A culture device for detection or enumeration of microorganisms, the device comprising an assay surface, the assay surface comprising liquid-retaining zones comprising a hydrophilic material and a land area comprising a hydrophobic material between the zones, at least one of the zones comprising an assay reagent.

40. The culture device of claim 39 wherein at least one of the liquid-retaining zones has a microvolume capacity of liquid retention.

41. The culture device of claim 39 wherein the device comprises about 10 to about 10,000 zones.

42. The culture device of claim 41 wherein the device comprises about 400 to about 600 zones.

43. The culture device of claim 39 wherein each zone has a liquid retention capacity of about 0.01 to about 25 microliters.

44. The culture device of claim 43 wherein each zone has a liquid retention capacity of about 1 to about 2 microliters.

45. The culture device of claim 39 wherein the assay reagent comprises a nutrient medium.

46. The culture device of claim 39 wherein the assay reagent comprises an indicator substance.

47. The culture device of claim 46 wherein the indicator substance is a chromogenic indicator, a fluorescent indicator, a luminescent indicator or an electrochemical indicator.

48. The culture device of claim 39 wherein the liquid-retaining zones comprise microvolume wells.

49. The culture device of claim 39 wherein the land area comprises a treated nanostructured film.

50. The culture device of claim 39 wherein the liquid-retaining zones comprise hydrophilic fiber material projecting from the hydrophobic material.

51. The culture device of claim 50 wherein at least one zone comprises a hydrophilic absorbent disc.

52. The culture device of claim 50 wherein at least one zone comprises protruding hydrophilic nonwoven fiber loop material.

53. The culture device of claim 39 wherein the culture device comprises a plurality of at least two sets of liquid-retaining zones, each set having zones of uniform size, the sets varying in liquid retention capacity.

54. A method for partitioning an aqueous liquid sample into discrete volumes to conduct a most probable number analysis, comprising:

a) providing a device for culturing a microorganism, the device including an assay surface, the assay surface comprising liquid-retaining zones comprising a hydrophilic material and a land area comprising a hydrophobic material between the zones;

b) contacting a liquid sample with the assay surface such that the liquid sample partitions into the liquid-retaining zones;

c) incubating the device;

d) detecting a signal that indicates microorganisms are growing in a zone; and

e) conducting a most probable number analysis based on the number of zones wherein a signal is detected.