Sample processing droplet actuator, system and method

• US 8,845,872 B2
• Filed: 09/30/2011
• Issued: 09/30/2014
• 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., Pamula, Vamsee K., Srinivasan, Vijay
• Primary Examiner(s): Dieterle, Jennifer
• Assistant Examiner(s): Kaur, Gurpreet
• Application Number:
  US13/249,425
• Publication Number:
  US 20120018306A1
• Time in Patent Office: 1,096 Days
• Field of Search: 204/600, 204/660, 204/663, 436/150, 422/504, 422/82, 422/52, 422/53
• US Class Current: 204/450
• 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

Sample processing droplet actuators, systems and methods are provided. According to one embodiment, a stamping device including a droplet microactuator is provided and includes: (a) a first plate including a path or network of control electrodes for transporting droplets on a surface thereof; (b) a second plate mounted in a substantially parallel orientation with respect to the first plate providing an interior volume between the plates, the second plate including one or more stamping ports for transporting some portion or all of a droplet from the interior volume to an exterior location; (c) a port for introducing fluid into the interior volume between the plates; and (d) a path or network of reference electrodes corresponding to the path or network of control electrodes. Associated systems and methods including the stamping device are also provided.

Claims

1. A method of dispensing at least one sample droplet from a parent droplet, the method comprising:

(a) providing a droplet actuator comprising substrates arranged to form a gap, wherein;

(i) the gap comprises;

(1) the parent droplet comprising a protein in a concentration exceeding 10 mg/mL;

(2) an oil filler fluid surrounding the parent droplet, the filler fluid comprising a, surfactant in an amount selected to enhance droplet operations; and

(ii) one or more of the substrates comprise electrodes arranged for conducting droplet operations in the gap by electrowetting using the parent droplet; and

(b) using the electrodes to conduct the droplet operations comprising dispensing at least one sample droplet from the parent droplet.

2. The method of claim 1 wherein the protein concentration exceeds 50 mg/mL.

3. The method of claim 1 wherein the protein concentration exceeds 100 mg/mL.

4. The method of claim 1 wherein the oil flier fluid comprises the surfactant in an amount which ranges from 0.001 to 10% w/w.

5. The method of claim 1 wherein the oil filler fluid comprises the surfactant in an amount which ranges from 0.001 to 1% w/w.

6. The method of claim 1 wherein the oil filler fluid comprises the surfactant in an amount which ranges from 0.001 to 0.1% w/w.

7. The method of claim 1 wherein the parent droplet has a volume which does not exceed 100 nL.

8. The method of claim 1 wherein the parent droplet has a volume which does not exceed 50 nL.

9. The method of claim 1 wherein the parent droplet has a volume which does not exceed 20 nL.

10. The method of claim 1 wherein the parent droplet has a volume which does not exceed 10 nL.

11. The method of claim 1 further comprising repeating the dispensing step to produce a set of sample droplets with <

2% CV.

12. The method of claim 1 wherein an enhancement in droplet operations comprises a reduction in protein contamination of droplet microactuator surfaces.

13. The method of claim 1 wherein the surfactant enables the dispensing of the at least one sample droplet.

14. The method of claim 1 wherein the oil filler fluid comprising the surfactant increases compatible protein concentration by more than 3 orders of magnitude relative to oil filler fluid in the absence of the surfactant.

15. The method of claim 1 wherein the surfactant increases the concentration of protein that can be reliably dispensed on the droplet actuator.

16. The method of claim 1 wherein the oil filler fluid comprises silicone oil.

17. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected from on-actuator sample reservoirs.

18. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected using from 1 mV to 50 kV.

19. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected using from 1V to 10 kV.

20. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected using from 5V to 1000V.

21. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected using from 10V to 300V.

22. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected using AC.

23. The method of claim 1 wherein the dispensing of the at least one sample droplet is effected using DC.