Droplet-based particle sorting

• US 8,883,513 B2
• Filed: 02/18/2011
• Issued: 11/11/2014
• Est. Priority Date: 04/18/2006
• Status: Expired due to Fees

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Pollack, Michael G., Pamula, Vamsee, Srinivasan, Vijay
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US13/030,418
• Publication Number:
  US 20110186433A1
• Time in Patent Office: 1,362 Days
• Field of Search: 422400-404, 422 63- 681, 422 81- 82, 422/82.01, 422/93, 422502-504, 422551-553, 436/180, 436/43, 436/174, 436/164, 436/158, 436/53, 204/450, 204600-604, 204/607, 204/608
• US Class Current: 436/164
• 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 method of providing a droplet comprising a single particle, the method comprising:

(a) providing a droplet comprising a suspension of particles on an electrowetting droplet actuator surrounded by an oil medium;

(b) dispensing from the droplet of (a) to provide a dispensed droplet, wherein the dispensing is electrowetting-mediated without the use of dielectrophoresis;

(c) determining whether the dispensed droplet on the electrowetting droplet actuator comprises a single particle; and

(d) sorting the dispensed droplet on the electrowetting droplet actuator based on the results of the determining step using electrowetting-mediated droplet operations.

2. The method of claim 1 further comprising repeating steps 23(b), 23(c), and 24(d).

3. The method of claim 1 wherein the dispensing is electrode-mediated.

4. The method of claim 1 wherein the dispensing is dielectrophoresis-mediated.

5. The method of claim 1 wherein the oil medium is substantially immiscible with the droplet.

6. The method of claim 1, further wherein the single particle comprises a target particle type, and wherein step 23(c) further comprises detecting the target particle type of the single particle in the dispensed droplet based on a property of and/or signal detected from the dispensed droplet.

7. The method of claim 1 further comprising combining a dispensed droplet comprising multiple particles with the suspension of particles.

8. The method of claim 1 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.

9. The method of claim 1 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.

10. The method of claim 1 further comprising transporting a dispensed droplet not comprising a single particle back to the suspension of particles using electrowetting-mediated droplet operations.

11. The method of claim 1 wherein a dispensed droplet lacking particles is discarded.

12. The method of claim 1 wherein single particle-containing droplets are transported to on-droplet microactuator reservoirs.

13. The method of claim 1 wherein additional droplet operations are conducted on a dispensed droplet comprising a single particle.

14. The method of claim 1 wherein a dispensed droplet comprising a single particle is further analyzed.

15. The method of claim 1 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.

16. The method of claim 15 wherein the detecting step is achieved using a sensor.

17. The method of claim 1 further comprising combining a droplet comprising a single particle with a droplet comprising a substance which may interact with the single particle.

18. The method of claim 17 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 method of claim 17 further comprising determining the presence or quantity of a substance indicative of the particle'"'"'s response to the substance.

20. The method of claim 19 wherein the determining step comprises conducting an assay comprising droplet-based steps.

21. The method of claim 17 wherein the particle comprises a T cell.

22. The method of claim 1 wherein step 23(c) is based on a signal detected within the dispensed droplet.

23. The method of claim 22 wherein the signal comprises a fluorescence from a molecule which binds to the particle.

24. The method of claim 22 wherein the signal comprises radioactive particles.

25. The method of claim 22 wherein the signal comprises a visual image.