Bead manipulation techniques

• US 9,205,433 B2
• Filed: 04/27/2015
• Issued: 12/08/2015
• Est. Priority Date: 04/13/2006
• Status: Active Grant

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Thwar, Prasanna, Pamula, Vamsee K., Sudarsan, Arjun, Sista, Ramakrishna
• Primary Examiner(s): Sines, Brian J
• Application Number:
  US14/697,058
• Publication Number:
  US 20150238975A1
• Time in Patent Office: 225 Days
• Field of Search: 422/50, 422/68.1, 422/82.01, 422/502, 422/503, 422/504, 436/43, 436/149, 436/150, 436/174, 436/179, 436/180
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• 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: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 2201/26 for use in medical applications
• CPC Class Codes: B03C 5/00 Separating dispersed particles from liquids by electrostatic effect flocculation or agglomeration of electric particles induced by electric field B01D21/0009; combined with centrifuges B04B5/10
• CPC Class Codes: B03C 5/005 Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength B03C5/02 takes precedence; electrophoresis B01D57/02
• CPC Class Codes: B03C 5/026 using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
• CPC Class Codes: B03C 7/026 using travelling or oscillating electric fields
• CPC Class Codes: G01N 33/54326 Magnetic particles

Abstract

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.

Claims

1. A droplet actuator comprising:

(a) a first substrate comprising;

(i) a droplet operations surface; and

(ii) electrodes arranged for conducting one or more electrowetting-mediated droplet operations on the surface; and

(b) one or more dielectrophoresis electrode configurations arranged for attracting and/or trapping one or more particles in a droplet situated on the droplet operations surface.

2. The droplet actuator of claim 1 wherein the droplet actuator comprises a second substrate separated from the droplet operations surface to form a droplet operations gap.

3. The droplet actuator of claim 2 wherein the one or more dielectrophoresis electrode configurations comprise at least one dielectrophoresis electrode configuration mounted on the second substrate.

4. The droplet actuator of claim 2 wherein the dielectrophoresis electrode configurations comprise at least one quadripole electrode configuration.

5. The droplet actuator of claim 4 wherein quadripole electrode configuration comprises four opposing triangular electrodes arranged to form a particle capture zone.

6. The droplet actuator of claim 5 wherein the four opposing triangular electrodes are symmetrical.

7. The droplet actuator of claim 5 wherein the four opposing triangular electrodes comprise one or more asymmetrical electrodes.

8. The droplet actuator of claim 4 wherein quadripole electrode configuration comprises four wires terminating at a particle capture zone.

9. The droplet actuator of claim 2 wherein the dielectrophoresis electrode configurations comprise at least one configuration comprising two electrodes the two electrodes comprising opposing fringed regions separated by a gap.

10. The droplet actuator of claim 2 wherein the dielectrophoresis electrode configurations comprise at least one configuration comprising multiple triangular electrodes arranged to form a particle trap zone.

11. The droplet actuator of claim 2 wherein the dielectrophoresis electrode configuration doubles as a droplet operations electrode.

12. The droplet actuator of claim 2 wherein the dielectrophoresis electrode configuration comprises a travelling wave configuration.

13. The droplet actuator of claim 2 wherein the gap comprises a filler fluid.

14. The droplet actuator of claim 13 the droplet is substantially surrounded by the filler fluid.

15. The droplet actuator of claim 13 wherein the filler fluid comprises an oil filler fluid.

16. The droplet actuator of claim 14 wherein the oil filler fluid comprises silicone oil.

17. A system comprising a processor coupled to the droplet actuator of claim 1 and programmed to control the droplet operations and/or the one or more dielectrophoresis electrodes.

18. A method of forming a droplet comprising one or more target particles, the method comprising:

(a) providing a droplet actuator, comprising;

(i) a first substrate comprising;

a droplet operations surface; and

electrodes arranged for conducting one or more electrowetting-mediated droplet operations on the surface; and

(ii) one or more dielectrophoresis electrode configurations arranged for attracting and/or trapping one or more particles in a droplet situated on the droplet operations surface;

(b) providing on the droplet operations surface a first droplet comprising a first concentration of particles;

(c) transporting the first droplet to an area of the droplet operations surface associated with one or more of the one or more dielectrophoresis electrode configurations;

(d) attracting and/or trapping one or more target particles in a region of the first droplet by activating one or more dielectrophoresis electrodes of one or more of the one or more dielectrophoresis electrode configurations; and

(e) forming a second droplet comprising the one or more target particles and a third droplet substantially lacking the target particles.

19. The method of claim 18 wherein forming the second and third droplet comprises splitting the first droplet using electrowetting-mediated droplet operations.

20. The method of claim 19 wherein the second droplet is retained in the area of the droplet operations surface associated with the one or more of the one or more dielectrophoresis electrode configurations, and the third droplet is transported away.

21. The method of claim 20 further comprising releasing the second droplet for transport and/or additional droplet operations.

22. The method of claim 21 wherein releasing the second droplet, comprises transporting a fourth droplet into contact with the second droplet to yield a combined droplet and deactivating the one or more dielectrophoresis electrodes.

23. The method of claim 21 wherein the second droplet comprising the one or more target particles has different dielectrophoresis properties relative to the third droplet substantially lacking the target particles.

24. The method of claim 20 wherein the second droplet is maintained in position by electrowetting-mediated droplet operations while the one or more target particles are attracted/trapped by the one or more dielectrophoresis electrodes.

25. A system comprising a processor programmed to execute the method of claim 18.