Manipulation of beads in droplets and methods for manipulating droplets

• US 9,494,498 B2
• Filed: 12/22/2015
• Issued: 11/15/2016
• 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: Pamula, Vamsee K., Sista, Ramakrishna, Srinivasan, Vijay, Pollack, Michael G., Eckhardt, Allen
• Primary Examiner(s): Sines, Brian J
• Application Number:
  US14/978,935
• Publication Number:
  US 20160109338A1
• Time in Patent Office: 329 Days
• Field of Search: 436/43, 436/149, 436/150, 436/174, 436/179, 436/180, 422/50, 422/68.1, 422/502, 422/503, 422/504, 422/521
• 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 dispensing a bead-containing sub-droplet, the method comprising:

(a) circulating magnetically responsive beads within a droplet on a droplet actuator, wherein circulating comprises;

(i) providing a droplet actuator, comprising;

(1) top and bottom substrates;

(2) 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

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

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

(iii) selectively altering at least one of the magnetic fields to circulate the magnetically responsive beads in the droplet; and

(b) using droplet operations to dispense a sub-droplet from 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.