METHODS AND DEVICES FOR SAMPLING FLOWABLE MATERIALS

• US 20090288710A1
• Filed: 09/12/2007
• Published: 11/26/2009
• Est. Priority Date: 09/13/2006
• Status: Active Grant

• Current Assignee: Centre National De La Recherche Scientifique, Institut Curie
• Original Assignee: Centre National De La Recherche Scientifique, Institut Curie
• Inventors: Chabert, Max, Viovy, Jean-Louis
• Granted Patent: US 8,741,661 B2
• Time in Patent Office: Days
• US Class Current: 137/1
• CPC Class Codes: B01F 33/3022 the components being formed by independent droplets which are alternated, the mixing of the components being achieved by diffusion between droplets
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/021 Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502769 characterised by multiphase flow arrangements
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 2035/00277 Special precautions to avoid contamination (e.g. enclosures, glove- boxes, sealed sample carriers, disposal of contaminated material)
• CPC Class Codes: G01N 35/08 using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: G01N 35/1009 Characterised by arrangements for controlling the aspiration or dispense of liquids
• CPC Class Codes: Y10T 137/0318 Processes
• CPC Class Codes: Y10T 137/85938 Non-valved flow dividers
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The present invention concern a method for sampling and mixing different flowable material materials from a plurality of wells in arbitrary order.

Claims

1. A method for manipulating at least one flowable material with a microsampler head having at least one flowable material inlet, said method comprising:

a) providing at least a first well with at least a first layer of a first flowable material and a second layer of a second flowable material immiscible with the first flowable material, located over said first layer, and at least a second well with at least a first layer of a first flowable material different from the first flowable material of said first well,

b) locating the at least one flowable material inlet of the microsampler head within the first layer of the first well,

c) sampling into the microsampler head a first volume of the first flowable material of the first well,

d) displacing the at least one flowable material inlet relative to the first well to locate the at least one flowable material inlet within the second layer,

e) sampling into the microsampler head a second volume of the second flowable material

f) displacing the at least one flowable material inlet relative to the first and second wells to locate the at least one flowable material inlet within the first layer of the second well,

g) sampling into the microsampler head a third volume of the first flowable material of the second well or dispensing into said second well a third volume of the first flowable material.

2. A method according to claim 1, wherein the first well is further provided with a third layer of a third flowable material located over the second layer and immiscible with the second flowable material.

3. A method according to claim 2, comprising displacing the at least one flowable material inlet relative to the first well after sampling the second volume of the second flowable material to move the at least one flowable material inlet from the second layer into the third layer of the first well.

4. A method according to claim 1, wherein the second well has over the first flowable material a second layer of a second flowable material immiscible with the first layer flowable material.

5. A method according to claim 4, wherein the second layers of the first and second wells are not in flowable material communication.

6. A method according to claim 4, comprising after sampling into the microsampler head the third volume of the first flowable material of the second well:

h) displacing the at least one flowable material inlet relative to the second well to locate the at least one flowable material inlet within the second layer of the second well,

i) sampling into the microsampler head a fourth volume of the second flowable material.

7. A method according to claim 6, wherein sampling at step h and sampling at step c are performed by identical or different ways selected among the active sampling and the passive sampling.

8. A method according to claim 4, wherein the second well comprises a third layer of a third flowable material immiscible with the second layer of flowable material.

9. A method according to claim 8, wherein the third layers of the first and second wells are in flowable material communication and consist of the same flowable material.

10. A method according to claim 9, comprising displacing laterally the at least one flowable material inlet within the third layer from over the first well to over the second well.

11. A method according to claim 4, wherein the second layer flowable material of the first and second wells comprise a same flowable material.

12. A method according to claim 2, wherein the third flowable material is less dense than the second flowable material.

13. A method according to claim 1, wherein the first flowable material of the first well comprises water.

14. A method according to claim 13, wherein the second flowable material of the first well comprises an organic solvent.

15. A method according to claim 2, wherein the third liquid of the first well comprises water.

16. A method according to claim 1, wherein the first flowable material of the first well comprises an organic solvent.

17. A method according to claim 16, wherein the second flowable material of the first well comprises water or a florinated liquid.

18. A method according to claim 2, wherein the third liquid comprises an organic solvent.

19. A method according to claim 1, wherein the first flowable material is less dense than the second flowable material.

20. A method according to claim 1, wherein the first flowable material of the first well comprises water and the first well comprises a first portion having an hydrophilic surface in contact with the first layer of the first flowable material.

21. A method according to claim 20, wherein the first well comprises a second portion having an hydrophobic surface.

22. A method according to claim 2, wherein the third flowable material is continuously renewed.

23. A method according to claim 1, wherein sampling is performed from more than 10 different wells.

24. A method according to claim 1, further comprising coalescing primary droplets of flowable material originating from the first and second wells.

25. A method according to claim 24, wherein the microsampler head comprises a first microchannel portion having a first cross-sectional area and a second microchannel portion having a second cross-sectional area larger that the first cross-sectional area, and wherein the coalescing occurs when entering the second microchannel portion.

26. A method according to claim 25, wherein the first microchannel portion is defined by a first duct force fitted into a second duct defining the second microchannel portion.

27. A method according to claim 25, wherein the at least one flowable material inlet is defined by a lower end of the first microchannel portion.

28. A method according to claim 1, wherein sampling at step c comprises creating a depression within the at least one flowable material inlet, as compared with the pressure at the top of said first layer.

29. A method according to claim 1, comprising sampling into the microsampler head a third volume of the first flowable material of the second well.

30. A method according to claim 1, comprising dispensing into the second well a third volume of the first flowable material.

31. A method according to claim 1, wherein the flowable material is a liquid.

36. A method for sampling a multiplicity of samples from a multiplicity of containers, comprising performing the method of claim 1.

37. A method for aliquoting at least one sample contained in a first container into a multiplicity of subsamples delivered to a multiplicity of second containers, comprising performing the method as defined in claim 1.

51. A method according to claim 1, whereas at least one element of a first flowable material with a volume less than 2 μ

l, is prepared as a droplet suspended in a second flowable material immiscible with said first flowable material.

32. A method for coalescing at least two primary droplets containing liquids, and respectively sampled into a microchannel from two different wells and thanks to a microsampler head having at least one flowable material inlet, said primary droplets being separated by a volume of an immiscible flowable material, wherein said primary droplets are displaced from a first portion of the microchannel having a first cross-sectional area to a second portion of the microchannel having a second cross-sectional area larger than said first cross-sectional area, and wherein the resulting droplet has a prolate shape in said second portion of the microchannel.

33. A method according to claim 32, wherein primary droplets are arranged in series, and wherein the primary droplets in a given series are coalesced together, but the resulting droplet from the coalescence of one series is not coalesced with any of the droplets resulting from the coalescence of primary droplets in other series.

34. A method for supplying an apparatus with flowable material samples, for example liquid samples, comprising:

thanks to first and second pumps, alternately drawing through a sample channel into first and second temporary storage channels droplets of samples in a carrier flowable material and alternately pushing droplets of samples in the carrier flowable material from the first and second buffer channels into an outlet channel, wherein when samples are drawn into the first buffer channel by the first pump samples are pushed by the second pump from the second buffer channel into the outlet channel and when samples are drawn into the second buffer channel by the second pump samples are pushed by the first pump from the first buffer channel into the outlet channel.

35. A method according to claim 34, wherein the first and second pumps are syringe pumps.

38. A sample-containing structure comprising a plurality of wells, each well of the plurality comprising a first portion being hydrophobic and a second portion being hydrophilic, at least one of the first and second portions being made of thermoplastic material.

39. A sampling structure according to claim 38, wherein the second portion comprises a coating of a fluorocarbon material adhering to an underlying portion made of said thermoplastic material or grafted onto it.

40. A sampling structure according to claim 38, wherein one of the first and second portions comprises a skirt and the other of the first and second portions comprises a well of a microtiter plate in which the skirt is engaged.

41. A sampling structure according to claim 40, comprising a cover plate comprising a plurality of skirts arranged to extend into corresponding wells of the microtiter plate.

42. A cover plate comprising:

an apertured support,

a plurality of skirts each extending below a corresponding aperture of the support and suitable for being inserted into a corresponding well of a microtiter plate on which the cover plate is placed.

43. A microsampler head comprising:

a first microchannel portion defining at least one flowable material inlet and having a first internal cross-sectional area,

a second microchannel portion in which the first duct portion opens out, the second microchannel portion having a second cross-sectional area larger than the first cross-sectional area,

wherein 1<

s₂/s₁≦

100 and s₂≦

10 mm².

44. A microsampler head according to claim 43, wherein the first microchannel portion is made of a first duct and the second microchannel portion is made of a second duct into which the first duct is inserted.

45. A multiplicity of droplets sequentially arranged in a microchannel into several series of droplets, wherein:

each droplet is essentially coaxial with the microchannel,

the shortest distance between two consecutive droplets in one series is never larger than a distance a, and the shortest distance along the microchannel'"'"'s axis between any two droplets belonging to two different series is never smaller than a.x, with x at least equal to 2,

and wherein at least two droplets in at least one of said series of droplets contain liquids and respectively sampled from two different wells and into said microchannel.

46. A device for flowable material sampling, for example liquid sampling, comprising a plurality of wells comprising at least a first well and a second well, each well comprising a first layer of a first flowable material covered by a second layer of a second flowable material immiscible with the first flowable material and a third layer of a third flowable material covering the second layer and immiscible with the second flowable material, wherein the first and second wells have different first flowable material and the same third flowable material and wherein the third layers of the first and second wells are in fluid communication.

47. A device according to claim 46, wherein said wells are arranged as an array compatible with the format of any of 48, 96, 384 or 1536-wells microtiter plates.

48. A device according to claim 46, wherein the first and second wells comprise first portions having an hydrophilic surface in contact with the respective first layers and wherein the first flowable material comprises water and are less dense than the respective second fluids.

49. A device according to claim 46, further comprising at least one microsampler head having at least one flowable material inlet and a drive mechanism for displacing the at least one flowable material inlet relative to the plurality of wells both vertically and horizontally.

50. A device according to claim 48, comprising a microchannel having an enlarged portion in which droplets originating respectively from the first and second wells coalesce.