Droplet-based nucleic acid amplification apparatus and system

• US 8,313,698 B2
• Filed: 12/06/2010
• Issued: 11/20/2012
• Est. Priority Date: 04/18/2006
• Status: Expired due to Fees

• Current Assignee: Duke University, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Duke University, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Pollack, Michael G., Paik, Philip Y., Pamula, Vamsee K.
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US12/960,583
• Publication Number:
  US 20110100823A1
• Time in Patent Office: 715 Days
• Field of Search: 422/50, 422/52, 422/68.1, 422 8205- 8208, 422/402, 422/417, 422/425, 422502-504, 422/518, 204/450, 204600-603
• US Class Current: 422/82.05
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1894 Cooling means; Cryo cooling
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• CPC Class Codes: B01L 2400/0427 Electrowetting
• 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: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: C12Q 2565/301 Pyrophosphate (PPi)
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

The present invention relates to a droplet-based nucleic acid amplification apparatus and system. According to one embodiment, a droplet microactuator is provided made using a first substrate including a fluorescing material and including a detection region for detecting a fluorescence signal from a droplet, which detection region is coated with a light absorbing, low fluorescence or non-fluorescing material.

Claims

1. A droplet microactuator, comprising:

a first substrate comprising a fluorescent material and comprising a detection region for detecting a fluorescence signal from a droplet, which detection region is coated with a light absorbing substantially non-fluorescent material;

electrodes arranged on the first substrate for conducting droplet operations; and

a processor electronically coupled to and configured for controlling the electrodes.

2. The droplet microactuator of claim 1 wherein the fluorescent material comprises a dielectric material.

3. The droplet microactuator of claim 1 wherein the coating comprises a metal coating.

4. The droplet microactuator of claim 1 wherein the coating comprises a copper coating.

5. The droplet microactuator of claim 1 wherein the coating comprises a black coating.

6. The droplet microactuator of claim 1 wherein the coating comprises a copper coating comprising a gold finish applied thereto.

7. The droplet microactuator of claim 1 further comprising a droplet on the substrate producing a fluorescent signal.

8. The droplet microactuator of claim 1 further comprising a droplet in the detection region, the droplet producing a fluorescent signal.

9. The droplet microactuator of claim 1 further comprising a second substrate spaced from and parallel to the first substrate, where a droplet is sandwiched between the first and second substrates.

10. The droplet microactuator of claim 9 wherein the first and/or second substrate comprise electrodes for conducting droplet operations.

11. The droplet microactuator of claim 10 wherein the electrodes are configured in an array.

12. The droplet microactuator of claim 10 wherein the electrodes comprise electrowetting electrodes configured for conducting electrowetting mediated droplet operation.

13. The droplet microactuator of claim 10 wherein the electrodes are arranged for conducting the droplet operations by dielectrophoresis.

14. The droplet microactuator of claim 1 wherein the electrodes are configured in an array.

15. The droplet microactuator of claim 1 wherein the electrodes comprise electrowetting electrodes configured for conducting electrowetting mediated droplet operation.

16. A droplet microactuator system comprising:

(a) a droplet actuator, comprising;

(i) a first substrate comprising a fluorescent material and comprising a detection region for detecting a fluorescence signal from a droplet, which detection region is coated with a light absorbing, substantially non-fluorescent material;

(ii) electrodes arranged on the first substrate for conducting droplet operations;

(iii) and a processor electronically coupled to and configured for controlling the electrodes; and

(b) a detector for detecting fluorescence from the droplet.

17. The droplet microactuator of claim 16 wherein the fluorescent material comprises a dielectric material.

18. The droplet microactuator system of claim 16 wherein the coating comprises a metal coating.

19. The droplet microactuator system of claim 16 wherein the coating comprises a copper coating.

20. The droplet microactuator system of claim 16 wherein the coating comprises a black coating.

21. The droplet microactuator system of claim 16 wherein the coating comprises a copper coating comprising a gold finish applied thereto.

22. The droplet microactuator system of claim 16 further comprising a droplet on the substrate producing a fluorescent signal.

23. The droplet microactuator system of claim 16 further comprising a droplet in the detection region, the droplet producing a fluorescent signal.

24. The droplet microactuator system of claim 16 further comprising a second substrate spaced from and parallel to the first substrate, where a droplet is sandwiched between the first and second substrates.

25. The droplet microactuator system of claim 24 wherein the first and/or second substrate comprise electrodes for conducting droplet operations.

26. The droplet microactuator system of claim 25 wherein the electrodes are configured in an array.

27. The droplet microactuator system of claim 25 wherein the electrodes comprise electrowetting electrodes configured for conducting electrowetting mediated droplet operation.

28. The droplet microactuator system of claim 25 wherein the electrodes are arranged for conducting the droplet operations by dielectrophoresis.

29. The droplet microactuator of claim 16 wherein the electrodes are configured in an array.

30. The droplet microactuator of claim 16 wherein the electrodes comprise electrowetting electrodes configured for conducting electrowetting mediated droplet operation.