Microfabricated crossflow devices and methods

• US 20020058332A1
• Filed: 09/14/2001
• Published: 05/16/2002
• Est. Priority Date: 09/15/2000
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Quake, Stephen R., Thorsen, Todd
• Granted Patent: US 7,294,503 B2
• Time in Patent Office: Days
• US Class Current: 435/288.3
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• 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 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0418 electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0454 radiation pressure, optical tweezers
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0484 Cantilevers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0638 membrane valves, flap valves
• 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 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• 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: C12M 1/34 : Measuring or testing with condition measuring or sensing means, e.g. colony counters
• CPC Class Codes: C12Q 1/68 : involving nucleic acids
• CPC Class Codes: G01N 15/01 : specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/1404 : Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1434 : Optical arrangements
• CPC Class Codes: G01N 15/1459 : the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1484 : microstructural devices
• CPC Class Codes: G01N 15/149 : specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 15/1492 : within droplets
• CPC Class Codes: G01N 2015/019 : Biological contaminants; Fouling
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1028 : Sorting particles
• CPC Class Codes: G01N 2015/1411 : Features of sheath fluids
• CPC Class Codes: G01N 2015/1481 : Optical analysis of particles within droplets sorting particles within droplets G01N15/1492
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• 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 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]

Abstract

A microfluidic device for analyzing and/or sorting biological materials (e.g., molecules such as polynucleotides and polypeptides, including proteins and enzymes; viruses and cells) and methods for its use are provided. The device and methods of the invention are useful for sorting particles, e.g. virions. The invention is also useful for high throughput screening, e.g. combinatorial screening. The microfluidic device comprises a main channel and an inlet region in communication with the main channel at a droplet extrusion region. Droplets of solution containing the biological material are deposited into the main channel through the droplet extrusion region. A fluid different from and incompatible with the solution containing the biological material flows through the main channel so that the droplets containing the biological material do not diffuse or mix. Biological material within the droplets can be analyzed and/or sorted by detecting a predetermined characteristic of the biological sample in each droplet and sorting the droplet accordingly.

Claims

1. A microfluidic device comprising

a main channel, and

at least one inlet region in communication with the main channel at a droplet extrusion region.

2. The device of claim 1 wherein a stream of an extrusion fluid passes through the main channel.

3. The device of claim 2 wherein a sample fluid, which is incompatible with the extrusion fluid, passes through the inlet region so that droplets of the second fluid are sheared into the main channel.

4. The device of claim 2 wherein the extrusion fluid is a non-polar solvent.

5. The device of claim 4 wherein the extrusion fluid is decane.

6. The device of claim 5 wherein the extrusion fluid is selected from the group consisting of tetradecane and hexadecane.

7. The device of claim 3 wherein the sample fluid comprises an aqueous solution.

8. The device of claim 7 wherein the aqueous solution comprises ultra pure water, TE buffer, phosphate buffer saline or acetate buffer.

9. The device of claim 3 wherein the sample fluid contains biological material.

10. The device of claim 9 wherein each of the droplets contains, on average, no more than one particle of the biological material.

11. The device of claim 10 wherein the biological material comprises viral particles and each of the droplets contains, on average, no more than one viral particle.

12. The device of claim 9 wherein the biological material comprises molecules of one or more enzymes.

13. The device of claim 9 wherein the biological material comprises molecules of one or more substrates.

14. The device of claim 9 wherein the biological material comprises one or more cells.

15. The device of claim 9 wherein the biological material comprises one or more viral particles.

16. The device of claim 1 wherein the droplet extrusion region comprises a T-shaped junction between the inlet region and the main channel.

17. The device of claim 1 further comprising:

a detection region within or coincident with at least a portion of the main channel at or downstream from the droplet extrusion region, and

a detector associated with the detection region.

18. The device of claim 17 wherein the detector is an optical detector.

19. The device of claim 17 further comprising:

a discrimination region downstream from the detection region, and

a flow control system responsive to the detector and adapted to direct droplets through the discrimination region into a branch channel.

20. The device of claim 1 wherein the main channel resides in a layer of elastomeric material.

21. The device of claim 20 wherein the elastomeric layer is adjacent to a substrate layer.

22. The device of claim 1, further comprising:

a first inlet region in communication with the main channel at a first droplet extrusion region; and

a second inlet region in communication with the main channel at a second droplet extrusion region.

23. The device of claim 22 wherein:

a first fluid containing a first biological material passes through the first inlet region, so that droplets of the fluid containing the first biological material are sheared into the main channel; and

a second fluid containing a second biological material passes through the second inlet region, so that droplets of the fluid containing the second biological material are sheared into the main channel.

24. The device of claim 23 wherein droplets containing the first biological material mix with droplets of the second biological material.

25. The device of claim 24 wherein:

the first biological material comprises an enzyme, and

the second biological material comprises a substrate to the enzyme.

26. The device of claim 24 wherein the first biological material interacts with the second biological material upon said mixing.

27. The device of claim 26 wherein the interaction produces a detectable signal.

29. The device of claim 28 wherein:

an extrusion fluid passes through the main channel, and

a sample fluid, which is incompatible with the first fluid and contains the biological material, passes through the inlet region, so that droplets of the second fluid and containing the biological material are sheared into the main channel.

30. The device of claim 29 wherein the flow control is adapted to direct the droplets into a branch channel.

31. The device of claim 29 wherein:

the extrusion fluid is a non-polar solvent, and

the sample fluid is an aqueous solution.

32. The device of claim 31 wherein the extrusion fluid is decane.

33. The device of claim 32 wherein the extrusion fluid is selected from the group consisting of tetradecane and hexadecane.

34. The device of claim 31 wherein the aqueous solution comprises ultra pure water, TE buffer, phosphate buffer saline, or acetate buffer.

35. The device of claim 29 wherein each of the droplets contains, on average, no more than one particle of the biological material.

36. The device of claim 35 wherein the biological material comprises viral particles and each of the droplets contains, on average, no more than one viral particle.

37. The device of claim 29 wherein the biological material comprises one or more viral particles.

39. The method of claim 38 wherein the biological material comprises one or more viral particles.

40. The method of claim 38 wherein:

the sample fluid is an aqueous solution; and

the extrusion fluid is a non-polar solvent.

41. The method of claim 40 wherein the aqueous solution comprises ultra pure water, TE buffer, phosphate buffer saline, or acetate buffer.

42. The method of claim 40 wherein the extrusion fluid is decane.

43. The method of claim 42 wherein the extrusion fluid is selected from the group consisting of tetradecane and hexadecane.

44. The method of claim 38 wherein each of the droplets contains, on average, no more than one particle of the biological material.

45. The method of claim 44 wherein the biological material comprises viral particles, and each of the droplets contains, on average, no more than one viral particle.

46. The device of claim 27, wherein the detectable signal is at least one of fluorescence and color change.

47. The device of claim 23, wherein the passage of each fluid through an inlet region is independently controlled.

48. The device of claim 23, wherein droplets of the first fluid are sheared into the main channel ahead of droplets of the second fluid.

49. The device of claim 23, wherein the passage of each fluid is controlled by at least one valve.

50. The device of claim 47, wherein the passage of each fluid is controlled by at least one valve.

51. The device of claim 19, wherein the flow control comprises the application of radiation pressure.

52. The device of claim 19, wherein the flow control comprises an optical beam.

53. The device of claim 3, wherein the sample fluid and the extrusion fluid have different refractive indexes.

54. The device of claim 3, wherein the refractive index of the sample fluid is lower than the refractive index of the extrusion fluid.

55. The device of claim 28, wherein the flow control comprises the application of radiation pressure.

56. The device of claim 28, wherein the flow control comprises an optical beam.

57. The device of claim 29, wherein the sample fluid and the extrusion fluid have different refractive indexes.

58. The device of claim 29, wherein the refractive index of the sample fluid is lower than the refractive index of the extrusion fluid.

59. The method of claim 38, wherein the refractive index of the extrusion fluid is different from the refractive index of the sample fluid, and wherein directing step comprises the application of radiation pressure.

60. The method of claim 59, wherein the application of radiation pressure comprises moving droplets with an optical beam.

28. A device for sorting biological material comprising:

(a) a microfabricated substrate having at least one main channel, an inlet region which meets the main channel at a droplet extrusion region, and at least two branch channels meeting at a junction downstream from the droplet extrusion region;

(b) a detection region within or coincident with at least a portion of the main channel and associated with a detector; and

(c) a flow control system responding to the detector and adapted to direct biological material into a branch channel.

38. A method for sorting biological material, said method comprising:

(a) furnishing an extrusion fluid to a main channel of a microfabricated substrate;

(b) providing droplets of a sample fluid containing the biological material to the main channel, wherein the sample fluid is incompatible with the extrusion fluid;

(c) interrogating the biological material in each droplet for a predetermined characteristic as it passes through a detection region associated with the main channel; and

(d) directing the flow of each droplet into a selected branch channel according to the results of the interrogation.