Particle capture devices and methods of use thereof

• US 9,201,060 B2
• Filed: 02/11/2009
• Issued: 12/01/2015
• Est. Priority Date: 02/11/2008
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology, Whitehead Institute
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Voldman, Joel, Skelley, Alison M., Kirak, Oktay, Jaenisch, Rudolf
• Primary Examiner(s): Flinders, Jeremy C
• Application Number:
  US12/867,031
• Publication Number:
  US 20110045994A1
• Time in Patent Office: 2,484 Days
• Field of Search: 506 33- 40, 435/288
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• 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 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/16 Surface properties and coatings
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• 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: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025

Abstract

The present invention provides a device and methods of use thereof in microscale particle capturing and particle pairing. This invention provides particle patterning device, which mechanically traps individual particles within first chambers of capture units, transfer the particles to second chambers of opposing capture units, and traps a second type of particle in the same second chamber. The device and methods allow for high yield assaying of trapped cells, high yield fusion of trapped, paired cells, for controlled binding of particles to cells and for specific chemical reactions between particle interfaces and particle contents. The device and method provide means of identification of the particle population and a facile route to particle collection.

Claims

1. A particle patterning device comprising:

a first substrate;

at least one capture unit positioned such that the top of said capture unit is adhered to or contiguous with said first substrate, said capture unit comprising;

a first chamber comprising at least one opening, sized to trap and accommodate a single particle; and

a second chamber comprising at least one opening, sized to trap and accommodate a single or more than a single particle;

wherein an opening of said first chamber faces a direction opposite to that of an opening of said second chamber, and said opening of said first chamber is contiguous with a side of said capture unit, said opening of said second chamber is contiguous with an opposite side of said capture unit;

a second substrate positioned proximally to or adhered to a bottom of said capture unit or a portion thereof;

a first conduit through which a first flow is induced in said device, such that said first flow is accommodated at least between said capture unit and said second substrate and particles positioned within said first or second chamber are subject to said first flow, wherein said first conduit, said capture unit, and said second substrate are configured to accommodate said first flow in a direction towards the opening of the first chamber; and

a second conduit through which a second flow is induced in a direction opposite to that of said first flow and towards the opening of the second chamber, such that said second flow is accommodated at least between said capture unit and said second substrate and particles positioned within the first or second chamber are subject to said second flow.

2. The particle patterning device of claim 1, further comprising at least one supporting structure positioned between said first and said second substrate, which aids in suspending said capture unit over said second substrate, at a height sufficient to accommodate fluid flow between said second substrate and said capture unit.

3. The particle patterning device of claim 2, wherein said at least one supporting structure is proximal to or contiguous with at least a portion of said capture unit.

4. The particle patterning device of claim 1, wherein said first chamber, said second chamber, or a combination thereof comprise at least one additional opening, which is sized such that said opening cannot trap or accommodate one or more particles.

5. The particle patterning device of claim 4, wherein said additional opening is positioned between said first and second chamber such that said opening facilitates fluid communication between said first and said second chamber.

6. The particle patterning device of claim 1, further comprising controllers to maintain desirable environmental conditions.

7. The particle patterning device of claim 6, wherein said controllers maintain a desired temperature, pH, CO₂ or Oxygen conditions, or a combination thereof.

8. The particle patterning device of claim 1, wherein said device is comprised of a transparent material.

9. The particle patterning device of claim 8, wherein said transparent material is pyrex, quartz, Polydimethylsiloxane (PDMS) or SU-8.

10. The particle patterning device of claim 1, further comprising electric connections between said first capture unit, said second capture unit or a combination thereof and a power supply.

11. The particle patterning device of claim 1, wherein said device is a microfluidic device.

12. The particle patterning device of claim 1, wherein said first chamber has a width ranging from between 5-500 μ

m and a depth ranging from between 5-500 μ

m.

13. The particle patterning device of claim 12, wherein said first chamber has a width of about 10 μ

m and a depth of about 5 μ

m.

14. The particle patterning device of claim 1, wherein said second chamber has a width ranging from between 5-500 μ

m and a depth ranging from between 5-500 μ

m.

15. The particle patterning device of claim 14, wherein said second chamber has a width of about 10 μ

m to about 35 μ

m and a depth of about 5.5 μ

m to about 50 μ

m.

16. The particle patterning device of claim 1, comprising an array of capture units.

17. The particle patterning device of claim 16, wherein said array is arranged in a row or column scheme or a combination thereof.

18. The particle patterning device of claim 17, wherein said scheme comprises a spacing between said row or column or combination thereof, said spacing approximate in width to a diameter of a particle being applied to said device.

19. The particle patterning device of claim 17, wherein said scheme comprises asymmetrically positioning a capture unit in a first row off-set from that of a capture unit in a preceding or subsequent row.

20. The particle patterning device of claim 19, wherein said off-set is half the center-to-center distance between two capture units.

21. An apparatus comprising the particle patterning device of claim 1.

22. The apparatus of claim 21, wherein an illumination source is coupled to said device.

23. The apparatus of claim 22, wherein the illumination source is a laser.

24. The apparatus of claim 22, wherein a beam splitter is employed with the use of said illumination source.

25. The apparatus of claim 21, further comprising a detector, which detects a change in a parameter in said device.

26. The apparatus of claim 25, wherein said detector comprises a camera, a computer, a luminometer, a spectrophotometer, or a combination thereof.

27. A method for patterning individual particles, said method comprising:

(i) applying a first liquid comprising a series of first particles to a particle patterning device under flow in a first direction, whereby an individual particle is accommodated within a first capture unit of said particle patterning device, said device comprising;

a first substrate;

at least one capture unit positioned such that the top of said capture unit is adhered to or contiguous with said first substrate, said capture unit comprising;

a first chamber comprising at least one opening, sized to trap and accommodate a single particle; and

a second chamber comprising at least one opening, sized to trap and accommodate a single or more than a single particle;

wherein an opening of said first chamber faces a direction opposite to that of an opening of said second chamber, and said opening of said first chamber is contiguous with a side of said capture unit, said opening of said second chamber is contiguous with an opposite side of said capture unit;

a second substrate positioned proximally to or adhered to a bottom of said capture unit or a portion thereof;

a first conduit through which a first flow is induced in said device, such that said first flow is accommodated at least between said capture unit and said second substrate and particles positioned within said first or second chamber are subject to said first flow, wherein said first conduit, said capture unit, and said second substrate are configured to accommodate said first flow in a direction towards the opening of the first chamber; and

a second conduit through which a second flow is induced in a direction opposite to that of said first flow and towards the opening of the second chamber, such that said second flow is accommodated at least between said capture unit and said second substrate and particles positioned within the first or second chamber are subject to said second flow; and

(ii) applying a second liquid to said device under flow in a second direction, wherein said individual particle moves from said first chamber of said first capture unit to a second chamber in a second capture unit.

28. The method of claim 27, wherein said particle patterning device comprises an array of capture units positioned in rows or columns or a combination thereof.

29. The method of claim 27, further comprising the step of applying a third liquid comprising a series of second particles to said device under flow in said second direction, whereby said series of second particles are patterned in said second chamber in a said second capture units with said series of first particles.

30. The method of claim 27, further comprising the step of applying a third liquid to said device under flow in said first direction, whereby said individual particle moves back to said first chamber in said first capture unit.

31. The method of claim 29, wherein said series of first particles, second particles or a combination thereof are cells, vesicles, or microspheres, or a combination thereof.

32. The method of claim 31, wherein said cells are mammalian cells, bacterial cells, parasitic cells, yeast cells or a combination thereof.

33. The method of claim 31, wherein said series of first particles and said series of second particles comprise cells of a similar cell type or are of, or isolated from, the same organism.

34. The method of claim 31, wherein said series of first particles and said series second particles comprise cells of a different cell type or are of, or isolated from, a different organism.

35. The method of claim 31, wherein said cells comprise a vector, which optionally comprises a reporter.

36. The method of claim 31, wherein said method is a fusion method between said cells, vesicles, or microspheres, or a combination thereof.

37. The method of claim 27, wherein said method further comprises applying a reagent to said device under flow in said second direction.

38. The method of claim 37, wherein said reagent is for detection, assay or a combination thereof of said series of first particles, second particles or a combination thereof.

39. The method of claim 37, wherein said reagent comprises a detectable marker.

40. The method of claim 29, wherein said series of first particles and said series of second particles are of the same size.

41. The method of claim 29, wherein said series of first particles and said series of second particles differ in size, composition, or a combination thereof.

42. The method of claim 29, further comprising the step of observing said series of first particles and said series of second particles over a course of time.

43. The method of claim 27, wherein said device is maintained under controlled temperature, pH, CO₂ or Oxygen conditions, or a combination thereof.

44. A method of contacting individual particles, said method comprising:

(i) applying a first liquid comprising a series of first particles to a particle patterning device under flow in a first direction, whereby an individual particle is accommodated within a first chamber in a first capture unit of said particle patterning device, said device comprising;

a first substrate;

at least one capture unit positioned such that the top of said capture unit is adhered to or contiguous with said first substrate, said capture unit comprising;

a first chamber comprising at least one opening, sized to trap and accommodate a single particle; and

a second chamber comprising at least one opening, sized to trap and accommodate a single or more than a single particle;

wherein an opening of said first chamber faces a direction opposite to that of an opening of said second chamber, and said opening of said first chamber is contiguous with a side of said capture unit, said opening of said second chamber is contiguous with an opposite side of said capture unit;

a second substrate positioned proximally to or adhered to a bottom of said capture unit or a portion thereof;

a first conduit through which a first flow is induced in said device, such that first flow is accommodated at least between said capture unit and said second substrate and particles positioned within said first or second chamber are subject to said first flow, wherein said first conduit, said capture unit, and said second substrate are configured to accommodate said first flow in a direction towards the opening of the first chamber such that a single particle introduced under flow is trapped within the first chambers; and

a second conduit through which a second flow is induced in a direction opposite to that of said first flow and towards the opening of the second chamber, such that said second flow is accommodated at least between said capture unit and said second substrate and particles positioned within the first or second chamber are subject to said second flow; and

(ii) applying flow in a second direction, wherein said individual particle moves from said first chamber of said first capture unit to a second chamber in a second capture unit; and

(iii) applying a second liquid comprising a series of second particles to said device in said second direction,

whereby said second particles positioned within said second chamber are brought into contact with said series of first particles, thereby being a method of contacting individual particles.

45. The method of claim 44, wherein bringing said first and second series of particles into contact results in a transfer of a material from one particle to another.

46. The method of claim 44, wherein said material comprise a biological material.

47. The method of claim 46, wherein said material is a nucleic acid or polypeptide.

48. The method of claim 44, wherein said particle is a cell.

49. The method of claim 48, wherein said method results in genetic transformation of said cell.

50. The method of claim 44, wherein said first or second series of particles or a combination thereof are cells.

51. The method of claim 50, wherein said method comprises a method of cell fusion.

52. The method of claim 50, wherein said method comprises a method of single cell infection.

53. The method of claim 50, wherein said method comprises a method of phagocytosis.

54. The method of claim 44, wherein bringing said first and said second series of particles into contact results in a measurable energy transfer.

55. The method of claim 54, wherein said energy transfer is fluorescent resonance energy transfer (FRET).

56. The method of claim 44, wherein said method comprises a screen for molecular interactions.