Manipulation of beads in droplets and methods for manipulating droplets

• US 9,377,455 B2
• Filed: 06/22/2015
• Issued: 06/28/2016
• 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: Sista, Ramakrishna, Pamula, Vamsee K., Srinivasan, Vijay, Pollack, Michael G., Eckhardt, Allen
• Primary Examiner(s): Sines, Brian J
• Application Number:
  US14/746,276
• Publication Number:
  US 20150285792A1
• Time in Patent Office: 372 Days
• Field of Search: 422/50, 422/68.1, 422/82.01, 422/82.02, 422/100, 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: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01F 33/3031 using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
• CPC Class Codes: B01F 33/451 wherein the mixture is directly exposed to an electromagnetic field without use of a stirrer, e.g. for material comprising ferromagnetic particles or for molten metal
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/06 Auxiliary integrated devices, integrated components
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• 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 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 2400/043 : magnetic forces
• CPC Class Codes: B01L 2400/0436 : acoustic forces, e.g. surface acoustic waves [SAW]
• CPC Class Codes: B01L 2400/0442 : thermal energy, e.g. vaporisation, bubble jet
• 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: B01L 3/502792 : for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: G01N 1/38 : Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 2001/386 : Other diluting or mixing processes
• CPC Class Codes: G01N 2035/1046 : Levitated, suspended drops
• CPC Class Codes: G01N 33/54326 : Magnetic particles
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25625 : Dilution
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The invention provides a method of circulating magnetically responsive beads within a droplet in a droplet actuator. The invention also provides methods for splitting droplets. The invention, in one embodiment, makes use of a droplet actuator with top and bottom substrates, a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable, and a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces. A droplet is positioned between the top and bottom surfaces and at least one of the magnetic fields is selectively altered.

Claims

1. A method of circulating magnetically responsive beads within a droplet on a droplet actuator, the method comprising:

(a) providing a droplet actuator, comprising;

(i) top and bottom substrates;

(ii) a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable; and

(iii) a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces;

(b) positioning the droplet comprising magnetically responsive beads between the top and bottom substrates in a gap that is within the magnetic fields; and

(c) selectively altering at least one of the magnetic fields to circulate the magnetically responsive beads in the droplet.

2. The method of claim 1 wherein altering at least one of the magnetic fields further comprises activating an electromagnet proximate at least one of the top and bottom surfaces.

3. The method of claim 1 wherein altering at least one of the magnetic fields further comprises activating and deactivating the magnetic fields in a pattern which causes the beads to circulate within the droplet.

4. The method of claim 1 wherein altering at least one of the magnetic fields further comprises activating an electromagnet proximate only one of the top and bottom surfaces.

5. The method of claim 1 further comprising selectively activating at least one of a plurality of electromagnets positioned proximate the top surface.

6. The method of claim 1 further comprising selectively deactivating at least one of a plurality of electromagnets positioned proximate the top surface.

7. The method of claim 1 further comprising selectively activating at least one of a plurality of electromagnets positioned proximate the bottom surface.

8. The method of claim 1 further comprising selectively deactivating at least one of a plurality of electromagnets positioned proximate the bottom surface.

9. The method of claim 1 wherein altering at least one of the magnetic fields further comprises physically altering a position of a magnet emanating the at least one magnetic field.

10. A method of circulating magnetically responsive beads within a droplet on a droplet actuator, the method comprising:

(a) providing a droplet actuator, comprising;

(i) top and bottom substrates;

(ii) a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable; and

(iii) a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces;

(b) positioning the droplet comprising magnetically responsive beads between the top and bottom substrates in a gap that is within the magnetic fields; and

(c) selectively altering at least one of the magnetic fields to circulate the magnetically responsive beads in the droplet without splitting, merging or transporting the droplet.

11. The method of claim 10 wherein altering at least one of the magnetic fields further comprises activating an electromagnet proximate at least one of the top and bottom surfaces.

12. The method of claim 10 wherein altering at least one of the magnetic fields further comprises activating an electromagnet proximate only one of the top and bottom surfaces.

13. The method of claim 10 further comprising selectively activating at least one of a plurality of electromagnets positioned proximate the top surface.

14. The method of claim 10 further comprising selectively deactivating at least one of a plurality of electromagnets positioned proximate the top surface.

15. The method of claim 10 further comprising selectively activating at least one of a plurality of electromagnets positioned proximate the bottom surface.

16. The method of claim 10 further comprising selectively deactivating at least one of a plurality of electromagnets positioned proximate the bottom surface.

17. The method of claim 10 wherein altering at least one of the magnetic fields further comprises physically altering a position of a magnet emanating the at least one magnetic field.