Multiwell droplet actuator, system and method

• US 7,998,436 B2
• Filed: 08/16/2007
• Issued: 08/16/2011
• 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., Srinivasan, Vijay, Pollack, Michael G.
• Primary Examiner(s): Gordon; Brian R
• Application Number:
  US11/839,774
• Publication Number:
  US 20080044893A1
• Time in Patent Office: 1,461 Days
• Field of Search: 422/99, 422/100, 422/500, 422502-504, 422/509, 422/63, 422 81- 82, 436/180
• US Class Current: 422/509
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/00587 High throughput processes
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/0893 having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 2400/043 magnetic forces
• 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: C07K 1/1136 by reversible modification of the secondary, tertiary or quarternary structure, e.g. using denaturating or stabilising agents
• CPC Class Codes: C30B 29/58 Macromolecular compounds
• CPC Class Codes: C30B 7/00 Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

Multiwell droplet actuators, systems and methods are provided. According to one embodiment, a substrate is provided and comprises: (a) one or more input ports for introduction of one or more reagents and/or samples; (b) a regular array of processing wells; and (c) a network of droplet transport pathways comprising pathways that provide direct or indirect droplet transport from each of the input ports to each of the one or more processing wells. Varying droplet actuators and systems related thereto are also provided.

Claims

1. An article of manufacture comprising:

(a) a substrate, a top plate mounted parallel to and separated from the substrate thereby providing a sealable interior volume between the substrate and the top plate, and one or more ports in the top plate arranged for introduction of one or more liquids into the interior volume and onto the substrate;

(b) an array of processing wells in the interior volume which are not in direct fluid communication with the one or more ports, each processing well defined by well walls, and wherein for each processing well the well walls comprise one or more openings in the well walls, the openings configured to enable liquid to be transported by electrowetting through the one or more openings and into each processing well, and wherein each processing well comprises one or more planar control electrodes in the substrate or top plate within the well walls; and

(c) a network of droplet transport pathways comprising pathways of electrodes arranged to provide direct or indirect electrowetting-mediated droplet transport from each of the ports through the openings in the walls and into each of the processing wells, wherein the one or more planar electrodes within the well walls of each processing well are arranged to operate in cooperation with the electrodes of the droplet transport pathways to conduct droplet operations.

2. The article of manufacture of claim 1 having footprint dimensions 85.48×

127.76 mm2.

3. The article of manufacture of claim 1 comprising 96 wells in an 8 by 12 format having a well-to-well pitch of 9 mm.

4. The article of manufacture of claim 1 wherein the network of droplet transport pathways interconnects the processing wells and is arranged such that a droplet extracted by electrowetting from any processing well can be transported via the network of droplet transport pathways to any other processing well via the network of droplet transport pathways.

5. The article of manufacture of claim 1 further comprising a means for localized heating of a processing well.

6. The article of manufacture of claim 1 further comprising at least one heating element arranged to provide localized heating of a region of the network of droplet transport pathways.

7. A system comprising:

(a) the article of manufacture of claim 6; and

(b) a processor electronically coupled to the article of manufacture and the at least one heating element, such that the processor controls the operation of the article of manufacture and the at least one heating element; and

wherein the processor is programmed to cause the article of manufacture to transport a droplet into proximity with the heating element causing heating of the droplet.

8. The system of claim 7 comprising two or more heating elements, wherein the processor is programmed to cause the article of manufacture to repetitively transport the droplet into proximity with one or more of the heating elements to subject the droplet to a heating and cooling cycle.

9. The system of claim 8 wherein the processor is programmed to control and cause the article of manufacture to execute a repetitive droplet heating protocol for amplification of DNA.

10. The article of manufacture of claim 6 comprising two or more heating elements spaced to maintain temperature zones on the substrate.

11. The system of claim 8 wherein the processor is programmed to control the temperature of the two or more heating elements, causing heating of the droplet but without causing significant vaporization of the droplet.

12. The article of manufacture of claim 1, wherein the pathways of electrodes establish multiple paths from each of the processing wells to each of the other processing wells.

13. The article of manufacture of claim 1, wherein the pathways of electrodes establish multiple intersecting paths from each of the processing wells to each of the other processing wells.

14. The article of manufacture of claim 1, wherein the pathways of electrodes establish multiple intersecting paths surrounding each of the processing wells.

15. The article of manufacture of claim 1 further comprising an input reservoir, wherein the one or more ports is arranged for introduction of one or more liquids into the interior volume and onto the substrate and is arranged for introduction of the one or more liquids into the input reservoir.

16. The article of manufacture of claim 1, wherein the planar electrodes are arranged together with the electrodes of the droplet transport pathways to conduct droplet dispensing operations from the processing wells.

17. The article of manufacture of claim 6 wherein the at least one heating element comprises a thin film heater.

18. The article of manufacture of claim 6 wherein the at least one heating element comprises a resistive trace heater.

19. The article of manufacture of claim 6 wherein the at least one heating element comprises a light heater.

20. The article of manufacture of claim 6 wherein the at least one heating element comprises a thermal heater.

21. The article of manufacture of claim 6 wherein the at least one heating element comprises apparatus for effecting Joule heating by passing electrical current through a droplet.

22. The article of manufacture of claim 6 wherein the at least one heating element comprises an infrared heater.

23. The article of manufacture of claim 6 wherein the interior volume is filled with an oil.