Techniques and droplet actuator designs for reducing bubble formation

• US 9,815,061 B2
• Filed: 11/20/2014
• Issued: 11/14/2017
• Est. Priority Date: 06/27/2012
• Status: Active Grant

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated), Illumina France Sarl (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated), Illumina France Sarl (Illumina Incorporated)
• Inventors: Delattre, Cyril, Rival, Arnaud, Srinivasan, Vijay
• Primary Examiner(s): Ball, J. Christopher
• Application Number:
  US14/549,113
• Publication Number:
  US 20150075986A1
• Time in Patent Office: 1,090 Days
• CPC Class Codes: B01L 2200/06 Fluid handling related problems
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/088 Channel loops
• 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 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 2400/0448 Marangoni flow; Thermocapillary effect
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• 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 3/502784 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 3/5029 using swabs
• CPC Class Codes: B01L 3/52 Containers specially adapted for storing or dispensing a reagent B01L3/02 takes precedence; containers for medical or pharmaceutical purposes A61J1/00; containers in general B65D; storing or dispensing test elements G01N33/4875; automated reagent dispensing G01N35/1002

Abstract

During droplet operations in a droplet actuator, bubbles often form in the filler fluid in the droplet operations gap and interrupt droplet operations. The present invention provides methods and systems for performing droplet operations on a droplet in a droplet actuator comprising maintaining substantially consistent contact between the droplet and an electrical ground while conducting multiple droplet operations on the droplet in the droplet operations gap and/or reducing the accumulation of electrical charges in the droplet operations gap during multiple droplet operations. The methods and systems reduce or eliminate bubble formation in the filler fluid of the droplet operations gap, thereby permitting completion of multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

Claims

1. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator, comprising;

a top substrate and a bottom substrate separated to form a droplet operations gap;

a conductive layer in contact with a surface of the top substrate, wherein the conductive layer provides a ground plane;

a dielectric layer in contact with the conductive layer, wherein an area of the dielectric layer has a sawtooth texture that extends into the droplet operations gap;

an arrangement of droplet operations electrodes in contact with the bottom substrate and arranged for conducting droplet operations thereon;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing a droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap,

wherein the droplet is transported through the filler fluid in the droplet operations gap; and

maintaining substantially consistent contact between the droplet and the sawtooth structure, and thus an electrical ground at the ground plane, while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

2. The method according to claim 1, further comprising heating the droplet in the droplet operations gap.

3. The method according to claim 1, further comprising heating the droplet in the droplet operations gap to at least sixty percent of a boiling point of the droplet.

4. The method according to claim 1, wherein conducting the multiple droplet operations comprises conducting at least 10 droplet operations without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

5. The method according to claim 1, further comprising heating the droplet in the droplet operations gap to at least sixty percent of a boiling point of the droplet, wherein conducting the multiple droplet operations comprises conducting at least 10 droplet operations without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

6. The method according to claim 5, wherein conducting the multiple droplet operations comprises conducting at least 100 droplet operations without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

7. The method according to claim 5, wherein conducting the multiple droplet operations comprises conducting at least 1,000 droplet operations without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

8. The method according to claim 5, wherein conducting the multiple droplet operations comprises conducting at least 100,000 droplet operations without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

9. The method according to claim 1, further comprising heating the droplet in the droplet operations gap to at least sixty percent of a boiling point of the droplet, wherein conducting the multiple droplet operations comprises completing an assay without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

10. The method according to claim 1, further comprising heating the droplet in the droplet operations gap to at least sixty percent of a boiling point of the droplet, wherein conducting the multiple droplet operations comprises completing multiple cycles of a polymerase chain reaction without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

11. The method according to claim 1, further comprising heating the droplet in the droplet operations gap to a minimum temperature of seventy five degrees Celsius, wherein conducting the multiple droplet operations comprises completing an assay without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

12. The method according to claim 1, further comprising heating the droplet in the droplet operations gap to within twenty degrees Celsius of a boiling point of the droplet, wherein conducting the multiple droplet operations comprises completing an assay without the interruption by the bubble formation in the filler fluid in the droplet operations gap.

13. The method according to claim 1, wherein the droplet comprises multiple droplets in the droplet operations gap, and wherein substantially consistent contact is maintained between multiple droplets and the electrical ground while conducting multiple droplet operations on the multiple droplets in the droplet operations gap.

14. The method according to claim 1, wherein the filler fluid is an electrically conductive filler fluid.

15. The method according to claim 1, further comprising merging the droplet with another droplet to maintain the substantially consistent contact with the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap.

16. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator comprising a top substrate and a bottom substrate separated to form a droplet operations gap, wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing the droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap, wherein the droplet is transported through the filler fluid in the droplet operations gap; and

adjusting a height of the droplet operations gap to maintain substantially consistent contact between the droplet and an electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

17. The method according to claim 16, further comprising grounding the top substrate of the droplet actuator to the electrical ground and maintaining substantially consistent contact between the droplet and the top substrate while conducting the multiple droplet operations on the droplet in the droplet operations gap.

18. The method according to claim 16, further comprising reducing the height of the droplet operations gap to maintain the substantially consistent contact between the droplet and the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap.

19. The method according to claim 16, further comprising adjusting the height of the droplet operations gap with a spring to maintain the substantially consistent contact between the droplet and electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap.

20. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator comprising a top substrate and a bottom substrate separated to form a droplet operations gap, wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing the droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap, wherein the droplet is transported through the filler fluid in the droplet operations gap; and

moving an electrical ground toward the droplet to maintain substantially consistent contact between the droplet and the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

21. The method according to claim 20 wherein:

the droplet actuator further comprises a plate and a set of adjustable ground probes that are electrically connected to the electrical ground;

the top substrate includes openings for respectively fitting the adjustable ground probes therethrough in a slidable fashion; and

the moving of the electrical ground toward the droplet is accomplished by pushing the plate toward the top substrate so that a tip of each of the adjustable ground probes extends through the openings and into the droplet operations gap.

22. The method according to claim 21 wherein the plate and the set of adjustable group probes are provided in a heated region of the droplet actuator.

23. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator comprising a top substrate and a bottom substrate separated to form a droplet operations gap, wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

bounding the droplet operations gap with a sidewall and an opposite sidewall to create a droplet operations channel;

arranging the droplet operations electrodes on the sidewall;

arranging one or more ground electrodes along the opposite sidewall;

connecting the one or more ground electrodes to an electrical ground;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing the droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap, wherein the droplet is transported through the filler fluid in the droplet operations gap; and

maintaining substantially consistent contact between the droplet and the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein the substantially consistent contact with the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap is unaffected by gravity;

and wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

24. The method according to claim 23, wherein the sidewall comprises a first rail and the opposite sidewall comprises a second rail, wherein the first rail and second rail are elongated three-dimensional (3D) structures that are arranged in parallel with each other.

25. The method according to claim 23, further comprising offsetting positions of the droplet operations electrodes and the position of the one or more ground electrodes.

26. The method according to claim 23, wherein the one or more ground electrodes are a continuous strip.

27. The method according to claim 23, further comprising oppositely arranging each droplet operations electrode to each of the one or more ground electrodes.

28. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator comprising a top substrate and a bottom substrate separated to form a droplet operations gap, wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

bounding the droplet operations gap with a sidewall and an opposite sidewall to create a droplet operations channel;

arranging the droplet operations electrodes on the sidewall;

arranging one or more ground electrodes along the bottom substrate;

connecting the one or more ground electrodes to an electrical ground;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing the droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap, wherein the droplet is transported through the filler fluid in the droplet operations gap; and

maintaining substantially consistent contact between the droplet and the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein the substantially consistent contact with the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap is unaffected by gravity;

and wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

29. The method according to claim 28, wherein the sidewall comprises a first rail and the opposite sidewall comprises a second rail, wherein the first rail and second rail are elongated three-dimensional (3D) structures that are arranged in parallel with each other.

30. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator comprising a top substrate and a bottom substrate separated to form a droplet operations gap, wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing the droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap, wherein the droplet is transported through the filler fluid in the droplet operations gap and wherein conducting the multiple droplet operations includes;

heating the droplet in the droplet operations gap;

applying a voltage to transport the droplet from an unactivated electrode to an activated electrode; and

reducing electrical charges in the droplet operations gap as the droplet is transported to the activated electrode by adjusting a height of the droplet operations gap; and

maintaining substantially consistent contact between the droplet and the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein bubble formation in the filler fluid in the droplet operations gap is reduced or eliminated;

and wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

31. The method according to claim 30, further comprising reducing the height of the droplet operations gap to reduce the electrical charges.

32. The method according to claim 30, further comprising texturing a surface of the top substrate to reduce the electrical charges.

33. A method of performing droplet operations on a droplet in a droplet actuator, comprising:

providing the droplet actuator comprising a top substrate and a bottom substrate separated to form a droplet operations gap, wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

filling the droplet operations gap of the droplet actuator with a filler fluid;

providing the droplet in the droplet operations gap;

conducting multiple droplet operations on the droplet in the droplet operations gap, wherein the droplet is transported through the filler fluid in the droplet operations gap and wherein conducting the multiple droplet operations includes;

heating the droplet in the droplet operations gap;

applying a voltage to transport the droplet from an unactivated electrode to an activated electrode; and

reducing discharge of electrical charges as the droplet is transported to the activated electrode by adjusting a height of the droplet operations gap; and

maintaining substantially consistent contact between the droplet and the electrical ground while conducting the multiple droplet operations on the droplet in the droplet operations gap;

wherein bubble formation in the filler fluid in the droplet operations gap is reduced or eliminated;

and wherein the substantially consistent contact between the droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.

34. The method according to claim 33, further comprising reducing the height of the droplet operations gap to reduce the discharge of electrical charges.

35. The method according to claim 33, further comprising texturing a surface of the top substrate of the droplet operations gap to reduce the discharge of electrical charges.

36. A system for performing droplet operations on a droplet in a droplet actuator, comprising a processor for executing code and a memory in communication with the processor, the system comprising code stored in the memory that causes the processor at least to:

provide the droplet in a droplet operations gap of the droplet actuator, wherein the droplet actuator comprises a top substrate and a bottom substrate separated to form the droplet operations gap, and wherein the droplet actuator further comprises an arrangement of droplet operations electrodes arranged for conducting droplet operations thereon;

fill the droplet operations gap of the droplet actuator with a filler fluid;

heat the droplet in a zone of the droplet operations gap to within twenty degrees Celsius of boiling to produce a heated droplet;

conduct multiple droplet operations on the heated droplet in the droplet operations gap, wherein the heated droplet is transported through the filler fluid in the zone of the droplet operations gap; and

adjust a height of the droplet operations gap to maintain substantially consistent contact between the heated droplet and an electrical ground while conducting the multiple droplet operations on the heated droplet in the zone of the droplet operations gap;

wherein the substantially consistent contact between the heated droplet and the electrical ground permits completion of the multiple droplet operations without interruption by bubble formation in the filler fluid in the zone of the droplet operations gap.