Droplet-Based Particle Sorting

• US 20110186433A1
• Filed: 02/18/2011
• Published: 08/04/2011
• Est. Priority Date: 04/18/2006
• Status: Active Grant

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Pamula, Vamsee K., Srinivasan, Vijay, Pollack, Michael G.
• Granted Patent: US 8,883,513 B2
• Time in Patent Office: Days
• US Class Current: 204/549
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• 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 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/089 Virtual walls for guiding liquids
• CPC Class Codes: B01L 2400/0427 Electrowetting
• 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 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• 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: 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 15/1031 by measuring electrical or magnetic effects
• CPC Class Codes: G01N 2015/019 Biological contaminants; Fouling
• CPC Class Codes: G01N 2015/1028 Sorting particles
• CPC Class Codes: G01N 2035/1034 Transferring microquantities of liquid
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 27/44717 Arrangements for investigating the separated zones, e.g. localising zones
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 35/10 Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The present invention relates to droplet-based particle sorting. According to one embodiment, a droplet microactuator is provided and includes: (a) a suspension of particles; and (b) electrodes arranged for conducting droplet operations using droplets comprising particles. A method of transporting a particle is also provided, wherein the method includes providing a droplet comprising the particle and transporting the droplet on a droplet microactuator.

Claims

1. A droplet microactuator comprising:

(a) a suspension of particles; and

(b) electrodes arranged for conducting droplet operations using droplets comprising particles.

2. The droplet microactuator of claim 1 wherein the electrodes are arranged for dispensing one or more droplets comprising one or more particles from the suspension of particles.

3. The droplet microactuator of claim 1 wherein the particles comprise biological cells.

4. The droplet microactuator of claim 1 wherein the particles comprise beads.

5. The droplet microactuator of claim 1 comprising a droplet comprising a single particle in the droplet.

9. A system comprising the droplet microactuator of claim 2 further comprising a sensor arranged for detecting a property of and/or a signal from a dispensed droplet.

10. The system of claim 9 wherein the system is programmed and configured with a sensor arranged for detecting the number of particles in the droplet based on the property of and/or signal from the dispensed droplet.

11. The system of claim 9 wherein the system is programmed and configured with a sensor arranged for detecting particle type of a particle in the droplet based on the property of and/or signal from the dispensed droplet.

12. The system of claim 11 wherein the particles are color coded.

13. The system of claim 11 wherein the particles are labeled.

14. The system of claim 11 wherein the particles comprise cells labelled with a labelled antibody.

15. The system of claim 11 wherein the particles comprise cells labelled with a fluorescing antibody.

16. The system of claim 9 wherein the system is programmed to combine a droplet comprising a single particle with a droplet comprising a substance which may interact with the single particle.

17. The system of claim 16 wherein the substance which may interact with the single particle is selected from the group consisting of:

beads, particles, microparticles, nanoparticles, drugs, proteins, peptides, antigens, toxins, and antibodies.

19. The system of claim 16 wherein the particle is a T particle.

20. The system of claim 9 comprising more than 50 droplets each comprising a predetermined number of particles.

21. The system of claim 9 comprising more than 250 droplets each comprising a predetermined number of particles.

22. The system of claim 9 comprising more than 1000 droplets each comprising a predetermined number of particles.

6. A droplet microactuator comprising a droplet comprising a single particle in the droplet.

7. The droplet microactuator of claim 6 wherein the droplet is surrounded by a filler fluid.

8. A method of transporting a particle comprising providing a droplet comprising the particle and transporting the droplet on a droplet microactuator.

18. The system of claim 18 wherein the system is further programmed to conduct an assay for the presence or quantity of a substance indicative of the particle'"'"'s response to the substance.

23. A method of providing a droplet comprising a single particle, the method comprising:

(a) providing a droplet comprising suspension of particles;

(b) dispensing a droplet from the droplet of (a) to provide a dispensed droplet; and

(c) determining whether the dispensed droplet comprises a single particle and/or a desired particle type.

24. The method of claim 23 further comprising repeating steps 23(b) and 23(c) until a predetermined number of droplets comprising single particles has been identified.

25. The method of claim 23 wherein the dispensing is electrode-mediated.

26. The method of claim 23 wherein the dispensing is electrowetting-mediated.

27. The method of claim 23 wherein the dispensing is dielectrophoresis-mediated.

28. The method of claim 23 wherein the droplet of 23(a) is provided surrounded by an oil medium.

29. The method of claim 28 wherein the oil medium is substantially immiscible with the droplet.

30. The method of claim 23 wherein step 23(c) is achieved by detecting the number of particles in the droplet based on a property of and/or signal from the dispensed droplet.

31. The method of claim 30 wherein the detecting step is achieved using a sensor.

32. The method of claim 23 wherein step 23(c) is achieved by detecting the particle type of a particle in the droplet based on a property of and/or signal from the dispensed droplet.

33. The method of claim 23 further comprising combining a droplet comprising a single particle with a droplet comprising a substance which may interact with the single particle.

34. The method of claim 33 wherein the substance which may interact with the single particle is selected from the group consisting of:

beads, particles, microparticles, nanoparticles, drugs, proteins, peptides, antigens, toxins, and antibodies.

35. The method of claim 33 further comprising determining the presence or quantity of a substance indicative of the particle'"'"'s response to the substance.

36. The method of claim 35 wherein the determining step comprises conducting an assay comprising droplet-based steps.

37. The method of claim 33 wherein the particle comprises a T cell.

38. The method of claim 23 further comprising combining a dispensed droplet comprising multiple particles with the suspension of particles.

39. The method of claim 23 further comprising:

(a) splitting a dispensed droplet comprising multiple particles to yield two or more daughter droplets; and

(b) analyzing one or more of the daughter droplets for the presence of a single particle.

40. The method of claim 23 further comprising:

(a) combining dispensed droplet comprising multiple particles a buffer droplet;

(b) splitting a dispensed droplet comprising multiple particles to yield two or more daughter droplets; and

(c) analyzing one or more of the daughter droplets for the presence of a single particle.

41. The method of claim 23 wherein a dispensed droplet lacking particles is transported back to the suspension of particles.

42. The method of claim 23 wherein a dispensed droplet lacking particles is discarded.

43. The method of claim 23 wherein sort decisions are based on a signal detected within single droplets.

44. The method of claim 43 wherein the signal comprises a fluorescence from a molecule which binds to the particle.

45. The method of claim 43 wherein the signal comprises radioactive particles.

46. The method of claim 43 wherein the signal comprises a visual image.

47. The method of claim 23 wherein single particle-containing droplets are transported to on-droplet microactuator reservoirs.

48. The method of claim 23 wherein droplet operations are conducted on single particle-containing droplets.

49. The method of claim 23 wherein single particle-containing droplets are analyzed.