Device and method for pressure-driven plug transport and reaction

• US 20050087122A1
• Filed: 01/26/2004
• Published: 04/28/2005
• Est. Priority Date: 05/09/2002
• Status: Active Grant

• Current Assignee: University of Chicago
• Original Assignee: University of Chicago
• Inventors: Ismagliov, Rustem F., Gerdts, Cory John, Zheng, Bo, Tice, Joshua David
• Granted Patent: US 7,901,939 B2
• Time in Patent Office: Days
• US Class Current: 117/2
• CPC Class Codes: B01D 2009/0086 Processes or apparatus therefor
• CPC Class Codes: B01D 9/0072 Crystallisation in microfluidic devices
• CPC Class Codes: B01F 25/433 Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
• CPC Class Codes: B01F 25/4331 Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01J 14/00 Chemical processes in general for reacting liquids with liquids; Apparatus specially adapted therefor B01J8/00, B01J19/08 take precedence
• 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 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00736 Non-biologic macromolecules, e.g. polymeric compounds
• CPC Class Codes: B01J 2219/00756 Compositions, e.g. coatings, crystals, formulations
• CPC Class Codes: B01J 2219/00783 Laminate assemblies, i.e. the reactor comprising a stack of plates
• CPC Class Codes: B01J 2219/00837 comprising coatings other than catalytically active coatings
• CPC Class Codes: B01J 2219/0086 Dimensions of the flow channels
• CPC Class Codes: B01J 2219/00869 Microreactors placed in parallel, on the same or on different supports
• CPC Class Codes: B01J 2219/00889 : Mixing micromixers B01F33/30
• CPC Class Codes: B01J 2219/00891 : Feeding or evacuation
• CPC Class Codes: B01J 2219/00894 : More than two inlets
• CPC Class Codes: B01J 2219/00903 : Segmented flow
• CPC Class Codes: B01J 2219/00975 : Ultraviolet light
• CPC Class Codes: B01J 2219/00977 : Infrared light
• CPC Class Codes: B01L 2200/0673 : Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/12 : Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0867 : Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• 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: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07K 14/43 : Sweetening agents, e.g. thaumatin, monellin
• CPC Class Codes: C12N 9/2462 : Lysozyme (3.2.1.17)
• CPC Class Codes: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Y 302/01017 : Lysozyme (3.2.1.17)
• CPC Class Codes: C30B 29/54 : Organic compounds
• CPC Class Codes: C30B 29/58 : Macromolecular compounds
• CPC Class Codes: C30B 7/14 : the crystallising materials being formed by chemical reactions in the solution
• CPC Class Codes: G01N 1/28 : Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• 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: Y10T 117/10 : Apparatus
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

The present invention provides microfabricated substrates and methods of conducting reactions within these substrates. The reactions occur in plugs transported in the flow of a carrier-fluid.

Claims

1. A method of crystallization, comprising the steps of:

a. introducing a first carrier-fluid into a first channel of a substrate;

b. introducing into a plug-forming region of the first channel;

1) a first plug-fluid comprising a precipitant;

2) a second plug-fluid comprising water; and

3) a third plug-fluid comprising a crystallization target;

where each of the plug-fluids comprises a solvent;

where pressure is applied when introducing each of the first, second and third plug-fluids, thereby resulting in the formation of a plurality of plugs in the first channel; and

where the crystallization target forms a crystal in at least one of the plugs.

2. The method of claim 1, further comprising the steps of:

a. introducing a second carrier-fluid into a second channel of the substrate;

b. introducing into a plug-forming region of the second channel;

1) a first plug-fluid comprising a precipitant;

2) a second plug-fluid comprising water; and

3) a third plug-fluid comprising a crystallization target;

where each of the plug-fluids comprises a solvent;

where pressure is applied when introducing each of the first, second and third plug-fluids, thereby resulting in the formation of a plurality of plugs in the channel;

where the first and second channels merge into a common downstream channel;

where at least one plug flowing from the first channel merges with a plug flowing from the second channel upon entry into the downstream channel.

4. The method of claim 1 or 3, where the crystallization target is a member of the group consisting of protein, peptide, polynucleotide, oligonucleotide, subcellular organelle, subcellular protein complex, drug, small molecule/biological macromolecule complex, virus, colloidal particle, nanoparticle and combinations thereof.

5. The method of claim 1 or 3, where the crystallization target is a protein.

6. The method of claim 1 or 3, where there is transfer of a solvent from one plug into another plug.

7. The method of claim 6, where the solvent is water.

8. The method of claim 1 or 3, where water is transported from one plug into another plug.

15. The method of claim 1 or 3, where each of the carrier-fluids and plug-fluids is introduced into the substrate according to a given flow rate under pressure, either directly into a channel or through an inlet feeding into a channel.

16. The method of claim 15, where the pressure is applied continuously.

17. The method of claim 15, where the pressure is halted after one or more plugs are formed.

18. The method of claim 15, where at least one of the flow rates is constant relative to one another flow rate.

19. The method of claim 15, where at least one of the flow rates is varied relative to one another flow rate.

20. The method of claim 15, where at least one of the plug-fluids is introduced into a carrier-fluid at a constant flow rate.

21. The method of claim 15, where at least one of the plug-fluids is introduced into a carrier-fluid at a variable flow rate.

22. The method of claim 15, where the flow rates are varied resulting in a plurality of plugs exhibiting a concentration gradient with regard to one or more of the plug-fluids.

23. The method of claim 1 or 3, where mixing of the plug-fluids within the plugs is increased by adding a turn to a channel.

26. The method of claim 1 or 3, where at least one of the plug-fluids comprises a salt.

35. The method of claim 1 or 3, where at least one of the carrier-fluids comprises an oil.

36. The method of claim 1 or 3, where at least one of the carrier-fluids comprises a fluorinated compound.

37. The method of claim 1 or 3, where at least one of the carrier-fluids comprises a surfactant.

38. The method of claim 1 or 3, where at least one of the carrier-fluids is permeable to a component in a plug.

39. The method of claim 38, where the component is selected from the group consisting of water, acid, base, buffer and solvent.

40. The method of claim 1 or 3, where at least one of the carrier-fluids is permeable to water.

45. The method of claim 1 or 3, where at least one of the carrier-fluids comprises poly-3,3,3-trifluoropropylmethylsiloxane (FMS-121).

46. The method of claim 1 or 3, where one of the carrier-fluids comprises perfluoroperhydrophenanthrene (PFP).

47. The method of claim 1 or 3, where one of the carrier-fluids comprises perfluoroperhydrophenanthrene (PFP) and 1H,1H,2H,2H-perfluorooctanol (PFO).

48. The method of claim 1 or 3, where the one of the carrier-fluids comprises 1H,1H,2H,2H-perfluorooctanol (PFO).

49. The method of claim 1 or 3, where at least one plug-fluid comprises a marker.

50. The method of claim 40, where the marker is an absorption dye or a fluorescent dye.

51. The method of claim 1 or 3, further comprising detecting the presence of a crystal in a plug.

52. The method of claim 1 or 3, further comprising analyzing the crystal in the capillary tube.

53. The method of claim 1 or 3, where a crystal is removed from a channel.

54. The method of claim 53, where the crystal is collected in a centrifuge tube.

55. The method of claim 53, where the crystal is collected in a sample tube.

56. The method of claim 53, where the crystal is collected in a micropipette.

57. The method of claim 53, where the crystal is collected in a capillary tube.

58. The method of claim 53, where the crystal is collected in a capillary tube and one or both ends of the capillary tube are sealed.

59. The method of claim 53, where the crystal is first collected in a capillary tube and then removed from the tube.

60. The method of claim 1 or 3, where the substrate comprises a material which allows water evaporation from the plugs.

61. The method of claim 1 or 3, where the substrate is soaked in water prior to crystal formation.

62. The method of claim 1 or 3, where the substrate is soaked in an aqueous salt solution prior to crystal formation.

63. The method of claim 1 or 3, where the substrate comprises a material which does not allow water evaporation from the plugs.

65. The method of claim 1, 3 or 64, where the second plug fluid further comprises a dehydration agent.

3. A method of crystallization, comprising the steps of:

a. introducing a first carrier-fluid into a first channel of a substrate;

b. introducing one or more first plug-fluids into a first plug-forming region of the first channel to form a plug of a first plug type;

where a plug of the first plug type comprises a precipitant and a crystallization target;

c. introducing one or more second plug-fluids into either one of;

1) the first plug-forming region;

2) a second plug-forming region within the first channel;

or

3) a second plug-forming region within a second channel;

where each of the plug-fluids comprises a solvent;

where the one or more second plug-fluids form a plug of a second plug type at either one of the first, second and third plug-forming regions;

where a plug of the second plug type comprises a precipitant;

where pressure is applied when introducing the one or more first plug fluids and the one or more second plug fluids;

where at least one or more plugs of the first plug type alternate in sequence with one or more plugs of the second plug type; and

where the crystallization target forms a crystal in at least one of the plugs.

9. The method of claim 3, where the one or more second plug-fluids is introduced into the first plug-forming region.

10. The method of claim 3, where the one or more second plug-fluids is introduced into the second plug-forming region.

11. The method of claim 3, where the one or more second plug-fluids is introduced into the second channel within the third plug-forming region.

12. The method of claim 3, where the first and second channels merge into a common downstream channel, such that one or more plugs flowing from the first channel alternate with one or more plugs of the second plug type flowing from the second channel upon entry into the downstream channel.

13. The method of claim 9, where a second carrier-fluid is introduced into the plug-forming region of the first channel so that it separates the one or more first plug-fluids from the one or more second plug-fluids prior to formation of the plug at the first plug-forming region.

14. The method of claim 13, where the second carrier-fluid is introduced through an inlet positioned between a first set of one or more inlets used for introducing one or more of the first plug-fluids and a second set of one or more inlets used for introducing one or more of the second plug-fluids.

24. The method of claim 3, where each of the carrier-fluids and plug-fluids is introduced into the substrate according to a given flow rate under pressure, either directly into a channel or through an inlet feeding into a channel;

and where one or more of the flow rates are varied so that the concentration of a first component in the plug of one plug-type provides a correlative and quantitative measure of a second component in an adjacent plug.

25. The method of claim 24, where the first component is a marker.

27. The method of claim 3, where the osmotic pressure in the plug of the first plug type at the first plug-forming region is lower than the osmotic pressure in the plug of the second plug type at the plug-forming region where the plug of the second type is formed.

28. The method of claim 3, where the osmotic pressure in the plug of the first type at the first plug-forming region is at least 2 times lower than the osmotic pressure in the plug of the second plug type at the plug-forming region where the plug of the second type is formed.

29. The method of claim 3, where the osmotic pressure in the plug of the first type at the first plug-forming region is at least 5 times lower than the osmotic pressure in the plug of the second plug type at the plug-forming region where the plug of the second type is formed.

30. The method of claim 3, where the osmotic pressure in the plug of the first type at the first plug-forming region is at least 10 times lower than the osmotic pressure in the plug of the second plug type at the plug-forming region where the plug of the second type is formed.

31. The method of claim 3, where introduction of the first plug-fluids and the second plug-fluids is regulated so that a plug flowing pattern is obtained in a common channel where at least 50% of the plugs of the first plug type are adjacent to plugs of the second plug type.

32. The method of claim 3, where introduction of the first plug-fluids and the second plug-fluids is regulated so that a plug flowing pattern is obtained in a common channel where at least 65% of the plugs of the first plug type are adjacent to plugs of the second plug type.

33. The method of claim 3, where introduction of the first plug-fluids and the second plug-fluids is regulated so that a plug flowing pattern is obtained in a common channel where at least 95% of the plugs of the first plug type are adjacent to plugs of the second plug type.

34. The method of claim 3, where a plug flowing pattern is obtained in which each of the plugs of the first plug type is adjacent to a plug of the second plug type.

41. The method of claim 3, where the first and second carrier-fluids are permeable to water.

42. The method of claim 3, where the first and second carrier-fluids are the same.

43. The method of claim 3, where the first and second carrier-fluids are the different.

44. The method of claim 3, where at least one of the carrier-fluids is impermeable to water.

64. A method of indexing a component in the plug of a microfluidic substrate, comprising the steps of:

a. introducing a first carrier-fluid into a first channel of a substrate;

b. introducing one or more first plug-fluids into a first plug-forming region of the first channel to form a plug of a first plug type;

where a cross section of a plug of the first plug type formed is substantially the same size as a cross section of the first channel;

c. introducing one or more second plug fluids into either one of;

1) the first plug-forming region;

2) the second plug-forming region within the first channel; and

3) a second plug-forming region within a second channel;

where the one or more second plug-fluids form a plug of a second plug type at either one of the first, second and third plug-forming regions;

where second plug comprises a marker;

where a cross section of plug of the second type formed is substantially the same size as a cross section of the channel in which it is formed;

where pressure is applied when introducing the one or more first plug fluids and the one or more second plug fluids, thereby inducing a flow of plugs in the substrate;

where at least one or more plugs of the first plug type alternate in sequence with one or more plugs of the second plug type;

where each of the carrier-fluids and plug-fluids is introduced into the substrate according to a given flow rate under pressure, either directly into a channel or through an inlet feeding into a channel; and

where one or more of the flow rates are regulated so that the marker provides a correlative and quantitative measure of a component in an adjacent plug.

66. A capillary tube comprising a plurality of plugs in an immiscible carrier-fluid, where a cross section of each plug is substantially the same size as a cross section of the capillary tube, said plurality of plugs comprising at least one plug having a crystallized target substance.

67. The capillary tube of claim 66, where the crystallization target is a member of the group consisting of protein, peptide, polynucleotide, oligonucleotide, subcellular organelle, subcellular protein complex, drug, small molecule/biological macromolecule complex, virus, colloidal particle, nanoparticle and combinations thereof.

68. The capillary tube of 66, where the crystallization target is a first protein.

69. The capillary tube of claim 68, where the first protein is bound to a synthetic molecule or a small molecule.

70. The capillary tube of claim 68, where the first protein is bound to a drug.

71. The capillary tube of claim 68, where the first protein is bound to a second protein or peptide.

72. The capillary tube of claim 68, where the first protein is bound to a member of the group consisting of oligonucleotides, carbohydrates and sugars.

73. The capillary tube of claim 68, where the carrier-fluid comprises an oil.

74. The capillary tube of claim 68, where the carrier-fluid comprises a fluorinated compound.