Droplet actuator and method

• US 9,861,986 B2
• Filed: 10/03/2014
• Issued: 01/09/2018
• Est. Priority Date: 05/03/2008
• Status: Active Grant

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Pollack, Michael G., Thwar, Prasanna, Pamula, Vamsee K., Srinivasan, Vijay, Eckhardt, Allen E., Shenderov, Alexander, Allen, Dwayne J.
• Primary Examiner(s): HIXSON, CHRISTOPHER
• Application Number:
  US14/506,265
• Publication Number:
  US 20150027889A1
• Time in Patent Office: 1,194 Days
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/022 Transponder chips
• CPC Class Codes: B01L 2300/023 Sending and receiving of information, e.g. using bluetooth
• CPC Class Codes: B01L 2300/024 Storing results with means integrated into the container
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• 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 2400/0415 electrical forces, e.g. electrokinetic
• 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 2400/0478 pistons
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 2400/0683 mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• 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: G01N 2035/1046 : Levitated, suspended drops
• CPC Class Codes: G01N 27/44704 : Details; Accessories
• CPC Class Codes: G01N 35/0098 : involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The invention provides droplet actuators and droplet actuator cassettes including reagent storage capabilities, as well as methods of making and using the droplet actuators and cassettes. The invention also provides continuous flow channel elements and techniques for using electrodes to manipulate droplets in flowing streams. The invention also discloses methods of separating compounds on a droplet actuator. Various other aspects of the invention are also disclosed.

Claims

1. A droplet actuator cartridge, comprising:

a droplet actuator portion, comprising;

two substrates separated by a droplet operations gap; and

one or more electrodes associated with one or both substrates and arranged for conducting one or more droplet operations in the droplet operations gap;

a reagent storage cassette portion, comprising;

a third substrate separated from one of the two substrates by a fluid channel;

one or more reservoirs formed in the third substrate, each of the one or more reservoirs having an opening to the fluid channel and comprising one or more liquids;

one or more plungers respectively insertable into the one or more reservoirs to force the one or more liquids through the opening and into the fluid channel; and

a removable protective film provided over the opening; and

a fluid path connecting the fluid channel to the droplet operations gap.

2. The droplet actuator cartridge of claim 1 wherein the one or more plungers force the one or more liquids from the one or more reservoirs into the fluid channel when depressed into the one or more reservoirs.

3. The droplet actuator cartridge of claim 2 comprising a series of the one or more reservoirs and a series of the one or more plungers, where each of the reservoirs is associated with a corresponding plunger arranged to force the one or more liquids from the reservoir into the fluid channel.

4. The droplet actuator cartridge of claim 3 wherein the one or more plungers are coupled to a common plunger depressor.

5. The droplet actuator cartridge of claim 2 wherein the removable protective film has physical characteristics, chemical characteristics, or both physical and chemical characteristics which permit the removable protective film to break upon application of pressure to the one or more liquids in the one or more reservoirs by depressing the one or more plungers.

6. The droplet actuator cartridge of claim 5 wherein the removable protective film is scored to facilitate breaking of the removable protective film upon application of pressure to the one or more liquids in the one or more reservoirs by depressing the one or more plungers.

7. The droplet actuator cartridge of claim 1 wherein the reagent storage cassette portion further comprises an awl, scribe or other puncturing device arranged to puncture the removable protective film and thereby permit the one or more liquids to flow into the fluid channel.

8. The droplet actuator cartridge of claim 7 wherein the awl, scribe or other puncturing device is slidably inserted within a slot in the one or more plungers.

9. The droplet actuator cartridge of claim 3, wherein upon depression of the one or more plungers, a series of droplets is forced into the fluid channel.

10. The droplet actuator cartridge of claim 9 wherein the fluid channel comprises a liquid filler fluid which is immiscible with the series of droplets.

11. The droplet actuator cartridge of claim 9 wherein the fluid channel is coupled to a pressure or vacuum source for flowing droplets through the fluid channel, through the fluid path and into the droplet operations gap.

12. The droplet actuator cartridge of claim 9 wherein the fluid channel is associated with one or more electrodes configured for transporting droplets through the fluid channel and into the droplet operations gap.

13. The droplet actuator cartridge of claim 1 further comprising a reagent on the one of the two substrates and in the fluid channel, wherein the reagent combines with the one or more liquids when the one or more liquids are forced into the fluid channel to yield a fully constituted reagent.

14. A droplet actuator, comprising:

two substrates separated by a droplet operations gap;

a droplet operations reservoir defined in the droplet operations gap;

an electrode associated with a first of the two substrates and arranged for conducting one or more droplet operations in the droplet operations reservoir;

a reagent reservoir defined in a second of the two substrates and aligned with the droplet operations reservoir;

a removable protective film separating the reagent reservoir and the droplet operations reservoir; and

a plunger associated with the reagent reservoir, the plunger being insertable into the reagent reservoir to force a liquid in the reagent reservoir directly into the droplet operations reservoir.

15. The droplet actuator of claim 14 wherein a top surface of the plunger is hydrophobic or is coated with a hydrophobic material.

16. A droplet actuator, comprising:

two substrates separated by a droplet operations gap;

a reservoir formed in a bottom of the two substrates;

an electrode positioned between the reservoir and the droplet operations gap, the electrode including an opening that provides a liquid path from the reservoir to the droplet operations gap;

a dielectric layer positioned on the electrode and at least in the opening;

a plunger extended partially into the reservoir, wherein the plunger is movable into a fully inserted position in the reservoir to force a liquid contained in the reservoir into the droplet operations gap when the liquid path is open; and

an awl inserted through an opening in the plunger, the awl being movable to a position that punctures the dielectric layer to open the liquid path and to a retracted position that permits the liquid to flow with reduced obstruction through the liquid path.

17. The droplet actuator of claim 16 wherein the electrode is a reservoir electrode, and wherein the droplet actuator further comprises droplet operations electrodes associated with the bottom of the two substrates and arranged for conducting one or more droplet operations in the droplet operations gap.

18. The droplet actuator of claim 17, further comprising a reference electrode associated with a top of the two substrates.

19. The droplet actuator of claim 16 wherein the electrode is a reference electrode and wherein the droplet actuator further comprises:

a reservoir electrode associated with a top of the two substrates and aligned with the reservoir; and

droplet operations electrodes associated with the top of the two substrates and arranged for conducting one or more droplet operations in the droplet operations gap.