Droplet-based particle sorting

• US 7,901,947 B2
• Filed: 12/15/2006
• Issued: 03/08/2011
• 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: Srinivasan, Vijay, Pollack, Michael G., Pamula, Vamsee K.
• Primary Examiner(s): Gordon; Brian R
• Application Number:
  US11/639,566
• Publication Number:
  US 20080053205A1
• Time in Patent Office: 1,544 Days
• Field of Search: 422/100, 422 63- 681, 436/180, 436/43, 436/174, 436/164, 204/450, 204/600
• US Class Current: 436/180
• 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 set of droplets, each comprising a single polymeric particle, the method comprising:

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

(b) dispensing a droplet from the droplet of (a) to provide dispensed droplets, wherein the dispensing is on the electrowetting droplet actuator and is electrowetting-mediated; and

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

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

2. The method of claim 1 wherein the method produces a predetermined number of droplets on the electrowetting droplet actuator, each droplet comprising a single polymeric particle.

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

4. The method of claim 1 wherein step (c) is achieved by detecting the number of polymeric particles in the droplet based on a property of and/or signal from the dispensed droplet.

5. The method of claim 4 wherein the detecting step is achieved using a sensor.

6. The method of claim 1 further comprising combining, on the electrowetting droplet actuator, a droplet comprising a single polymeric particle with a droplet comprising a substance which may interact with the single polymeric particle.

7. The method of claim 6 wherein the substance which may interact with the single polymeric particle is selected from the group consisting of:

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

8. The method of claim 6 further comprising determining the presence or quantity of a substance indicative of the polymeric particle'"'"'s interaction with the substance.

9. The method of claim 8 wherein the determining step comprises conducting an assay comprising droplet-based steps.

10. The method of claim 1 further comprising transporting a dispensed droplet not comprising a single polymeric particle back to the suspension of particles.

11. The method of claim 1 wherein droplet operations are conducted on single polymeric particle-containing droplets.

12. The method of claim 1 wherein single polymeric particle-containing droplets are analyzed.

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

(a) providing a droplet comprising a suspension of polymeric particles;

(b) dispensing from the droplet of (a) to provide dispensed droplets, wherein the dispensing comprises electrowetting-mediated droplet operations; and

(c) determining whether each dispensed droplet comprises a single polymeric particle;

(d) sorting the dispensed droplets based on results of the determining step, wherein the sorting comprises electrowetting-mediated droplet operations; and

(e) transporting back to the suspension of polymeric particles dispensed droplets not comprising a single polymeric particle, wherein the transporting comprises electrowetting-mediated droplet operations.

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

(a) providing a droplet comprising a suspension of polymeric particles;

(b) dispensing from the droplet of (a) to provide dispensed droplets, wherein the dispensing comprises electrowetting-mediated droplet operations; and

(c) determining whether each dispensed droplet comprises a single polymeric particle;

(d) sorting the dispensed droplets based on results of the determining step, wherein the sorting comprises electrowetting-mediated droplet operations;

(e) transporting back to the suspension of polymeric particles dispensed droplets not comprising a single polymeric particle, wherein the transporting comprises electrowetting-mediated droplet operations; and

(f) combining a droplet comprising a single polymeric particle with a droplet comprising a substance which may interact with the single polymeric particle, wherein the transporting comprises electrowetting-mediated droplet operations.

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

(a) providing on a droplet actuator without fixed microchannels a droplet comprising a suspension of polymeric particles;

(b) dispensing on the droplet actuator from the droplet of (a) to provide dispensed droplets on the droplet actuator, wherein the dispensing comprises electrowetting-mediated droplet operations; and

(c) determining whether each dispensed droplet comprises a single polymeric particle;

(d) sorting on the droplet actuator the dispensed droplets based on results of the determining step, wherein the sorting comprises electrowetting-mediated droplet operations; and

(e) transporting on the droplet actuator to the suspension of polymeric particles dispensed droplets not comprising a single polymeric particle, wherein the transporting comprises electrowetting-mediated droplet operations.