Method for carrying out a biochemical protocol in continuous flow in a microreactor

• US 6,977,145 B2
• Filed: 01/29/2001
• Issued: 12/20/2005
• Est. Priority Date: 07/28/1999
• Status: Expired due to Fees

• Current Assignee: Commissariat a L'Energie Atomique (French Alternative Energies and Atomic Energy Commission), Serono Genetics Institute S.A. (Merck & Co., Inc.)
• Original Assignee: Serono Genetics Institute S.A. (Merck & Co., Inc.)
• Inventors: Charles, Raymond, Clerc, Jean-Frederic, Vauchier, Claude, Peponnet, Christine, Sarrut, Nicolas, Fouillet, Yves, Claustre, Patricia
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): Tung, Joyce
• Application Number:
  US09/772,280
• Publication Number:
  US 20010041357A1
• Time in Patent Office: 1,786 Days
• Field of Search: 435/6, 435/91.2, 435/283.1, 435/286.5, 435/287.1, 435/288.4
• US Class Current: 435/6.19
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0845 Filaments, strings, fibres, i.e. not hollow
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1838 using fluid heat transfer medium
• CPC Class Codes: B01L 2300/1883 using thermal insulation
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• 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 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• 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 7/02 Water baths; Sand baths; Air baths
• 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 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: B01L 7/5255 : by moving sample containers
• CPC Class Codes: F28F 2260/02 : having microchannels
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis

Abstract

Devices and methods for carrying out a chemical or biochemical protocol are disclosed. In one embodiment, the chemical or biochemical protocol is performed by cycling at least one thermal transfer member between at least two temperatures while liquid samples on which the chemical or biochemical protocol is to be performed are continuously moving through at least one temperature regulated zone upon which the at least one thermal transfer member acts. In some embodiments, the device comprises a sample transport member that comprises liquid samples in sample receiving regions. The sample transport member moves the samples continuously through a temperature regulated zone which cycles between at least two temperatures while the liquid samples are moving through a temperature regulated zone on which at least one thermal transfer member acts. In some embodiments, the sample receiving regions comprise wells, hydrophillic films or hydrophillic filaments.

Claims

1. A method for carrying out a chemical or biochemical protocol comprising:

depositing liquid sample volumes into a plurality of sample receiving regions on at least one mobile sample transport member; and

moving the sample transport member along a pathway such that said sample receiving regions move through at least one temperature regulated zone upon which a thermal transfer member acts, wherein said thermal transport member cycles between at least two temperatures while said sample receiving regions are moving through said at least one temperature regulated zone; and

whwrein the protocol is carried out in an atmosphere sufficiently humid to reduce or prevent evaporation of the liquid sample volumes.

2. The method of claim 1 further comprising adding at least one reagent to the sample receiving regions while the sample receiving regions are moving along said pathway.

3. The method of claim 1 wherein the sample receiving regions comprise areas on a substrate.

4. The method of claim 3 wherein the areas on the substrate comprise wells.

5. The method of claim 4 wherein the sample receiving regions comprise a plate, having a plurality of wells therein, said wells having a thin film on their bottom surfaces.

6. The method of claim 3 wherein the substrate is a film.

7. The method of claim 6 wherein a surface of the film is sufficiently hydrophillic to allow adherence of individual liquid sample volumes in the form of droplets on the surface.

8. The method of claim 6 wherein said film comprises a matrix of hydrophobic areas and hydrophillic areas, said hydrophillic areas being sufficiently hydrophillic to allow adherence of individual liquid samples in the form of droplets on said hydrophillic areas.

9. The method of claim 3 wherein the substrate comprises a filament.

10. The method of claim 9 wherein the filament is sufficiently hydrophillic to allow adherence of individual liquid sample volumes in the form of droplets on the filament.

11. The method of claim 9 wherein the filament is conducting, and the droplets are heated by passing electric current through the filament.

12. The method of claim 1 wherein said sample transport member moves along said pathway continuously.

13. The method of claim 1 wherein said sample transport member moves said pathway in steps.

14. The method of claim 1 wherein said sample transport member is moved along said pathway by reels which frictionally engage the sample transport member.

15. The method of claim 1 wherein the sample receiving regions are covered by a non-miscible liquid in order to prevent evaporation of the liquid sample volumes.

16. The method of claim 1 wherein one of the at least two temperatures is about 50°

C., and another of the at least two temperatures is about 94°

C.

17. The method of claim 1 wherein said thermal transfer member cycles through said at least two temperatures a plurality of times while said sample receiving regions are moving through said at least one temperature regulated zone.

18. The method of claim 1 wherein said thermal transfer member cycles through said at least two temperatures from about 2 to about 35 times while said sample receiving regions are moving through said at least one temperature regulated zone.

19. The method of claim 1 wherein the protocol is carried out in only one apparatus.

20. The method of claim 1 wherein a plurality of sample receiving regions are processed in parallel in said at least one temperature regulated zone.

21. The method of claim 1 wherein said chemical or biochemical protocol comprises a nucleic acid amplification procedure.

22. The method of claim 21 wherein said chemical or biochemical protocol comprises a polymerase chain reaction.

23. The method of claim 21 wherein said chemical or biochemical protocol comprises determining the identity of at least one polymorphic nucleotide in the product of said nucleic amplification procedure.

26. The method according to claim 1, wherein said pathway is a channel.

27. The method according to claim 1, wherein said thermal transfer member is a metal bar in fluid communication with a plurality of water sources that provide water having said at least two temperatures.

28. The method according to claim 1, wherein said sample transport member is continuously flowing through said at least one temperature regulated zone.

31. The method according to claim 1, wherein said pathway is a microchannel.

24. A method for carrying out a chemical or biochemical protocol comprising:

depositing liquid sample volumes into a plurality of sample receiving regions on at least one mobile sample transport member; and

moving the sample transport member along a pathway such hat said sample receiving regions move through at least one temperature regulated zone upon which a thermal transfer member acts, wherein said thermal transport member cycles between at least two temperatures while said sample receiving regions are moving through said at least one temperature regulated zone;

wherein said sample transport member is moved along said pathway by reels which frictionally engage the sample transport member.

25. A method for carrying out a chemical or biochemical protocol comprising:

depositing liquid sample volumes into a plurality of sample receiving regions on at least one mobile sample transport member; and

moving the sample transport member along a pathway such that said sample receiving regions move through at least one temperature regulated zone upon which a thermal transfer member acts, wherein said thermal transport member cycles between at least two temperatures while said sample receiving regions are moving through said at least one temperature regulated zone; and

wherein the sample receiving regions are covered by a non-miscible liquid in order to prevent evaporation of the liquid sample volumes.

29. The method according to clam 3, wherein said substrate is a microfluidic substrate.

30. The method according to claim 29, wherein said microfluidic substrate comprises at least one microchannel.

32. The method according to claim 30, wherein said substrate is contacted with a thermal transfer member is a metal bar in fluid communication with a plurality of water sources that provide water having said at least two temperatures.

33. A method for carrying out a chemical or biochemical protocol comprising;

depositing liquid sample volumes into a plurality of sample receiving regions on at least one mobile sample transport member; and

moving the sample transport member along a pathway such that said sample receiving regions move through at least one temperature regulated zone upon which a thermal transfer member acts, wherein said thermal transport member cycles between at least two temperatures while said sample receiving regions are moving through said at least one temperature regulated zone.

34. A chemical or biochemical protocol comprising the steps:

a) providing at least one mobile sample transport member comprising at least one sample receiving region;

b) applying a sample to said at least one sample receiving region;

c) moving said at least one mobile transport member continuously along a pathway into a temperature regulated zone on which a thermal transfer member acts;

d) cycling said thermal transfer member between at least two temperatures.

35. The method of claim 34, wherein said thermal transfer member is cycled between at least two temperatures a plurality of times.

36. The method of claim 34, wherein said at least one mobile transport member is continuously moved along a pathway into another temperature regulated zone on which another thermal transfer member acts.

37. The method of claim 34, wherein said at least one mobile transport member is continuously moved along a pathway through a plurality of temperature regulated zones, each of said temperature regulated zones being acted upon by a thermal transfer member.

38. The method of claim 34, further comprising the step of adding reagent to said sample in said sample receiving region.

39. The method of claim 34, wherein said thermal transfer member is a metal bar in fluid communication with at least one heating, at least one cooling or at least one heating and at least one cooling reservoir containing a fluid.

40. The method of claim 39, wherein said fluid is a gas.

41. The method of claim 39, wherein said fluid is a liquid.

42. The method of claim 41, wherein said liquid is water.

43. The method of claim 35, wherein said thermal transfer member is a metal bar in fluid communication with at least one heating, at least one cooling or at least one heating and at least one cooling reservoir containing a fluid.

44. The method claim 43, wherein said fluid is a gas.

45. The method of claim 43, wherein said fluid is a liquid.

46. The method of claim 45, wherein said liquid is water.

47. The method of claim 36, wherein said thermal transfer member is a metal bar in fluid communication with at least one heating, at least one cooling or at least one heating and at least one cooling reservoir containing a fluid.

48. The method claim 47, wherein said fluid is a gas.

49. The method or claim 47, wherein said fluid is a liquid.

50. The method of claim 49, wherein said liquid is water.

51. The method of claim 37, wherein said thermal transfer member is a metal bar in fluid communication with at least one heating, at least one cooling or at least one heating and at least one cooling reservoir containing a fluid.

52. The method of claim 51, wherein said fluid is a gas.

53. The method of claim 51, wherein said fluid is a liquid.

54. The method of claim 53, wherein said liquid is water.