Method of Effecting Coagulation in a Droplet

• US 20110104725A1
• Filed: 05/04/2009
• Published: 05/05/2011
• Est. Priority Date: 05/02/2008
• Status: Abandoned Application

• Current Assignee: Children's Medical Center Corporation, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Children's Medical Center Corporation, Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Rouse, Jeremy, Eckhardt, Allen, Emani, Sitaram, Graham, Carrie, Pamula, Vamsee K.
• Application Number:
  US12/990,766
• Publication Number:
  US 20110104725A1
• Time in Patent Office: Days
• US Class Current: 435/7.92
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01F 33/3031 using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/089 Virtual walls for guiding liquids
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• 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 2400/046 Chemical or electrochemical formation of bubbles
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• 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: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors

Abstract

The invention provides techniques for coagulating blood on a droplet actuator. The invention also provides methods of manipulating the coagulated blood including a variety of droplet operations that may be conducted using the coagulated blood. Further, the invention provides a variety of assays that make use of the coagulated blood or various blood samples as input.

Claims

1. A method of effecting coagulation in source droplet, the method comprising:

(a) providing a liquid filler fluid;

(b) providing in the liquid filler fluid a source droplet comprising one or more coagulatable substances;

(c) treating in the liquid filler fluid the source droplet to effect coagulation of the one or more coagulatable substances to yield a coagulated droplet in the liquid filler fluid comprising a coagulated portion and supernatant.

2. The method of claim 1 wherein the source droplet comprises a biological fluid.

3. The method of claim 1 wherein the biological fluid comprises a blood sample.

4. The method of claim 3 wherein the blood sample comprises whole blood.

5. The method of claim 3 wherein the blood sample consists essentially of whole blood.

6. The method of claim 3 wherein the blood sample consists of whole blood.

7. The method of claim 3 wherein the blood sample comprises one or more natural blood components.

8. The method of claim 1 wherein the blood sample comprises one or more artificial blood components.

9. The method of claim 1 wherein the blood sample comprises one or more anticoagulants.

10. The method of claim 9 wherein the anticoagulant is selected from the group consisting of coumarines, vitamin K antagonists, acenocoumarol, phenprocoumon, brodifacoum, phenindione, heparins, low molecular weight heparin, synthetic pentasaccharide inhibitors of Factor Xa, and thrombin inhibitors.

11. The method of claim 8 wherein the one or more artificial blood components comprise one or more artificial platelet components.

12. The method of claim 8 wherein the one or more artificial blood components comprise one or more artificial oxygen carriers.

13. The method of claim 1 wherein the source droplet comprises a milk sample.

14. The method of claim 1 wherein the source droplet comprises a plant sample.

15. The method of claim 1 wherein the source droplet comprises coagulatable beads.

16. The method of claim 1 wherein step 1(c) comprises combining the sample droplet with a procoagulant droplet comprising a procoagulant.

17. The method of claim 1 wherein step 1(c) comprises contacting the sample droplet with a procoagulant.

18. The method of claim 1 wherein step 1(c) comprises incubating the sample droplet for a period of time sufficient to permit coagulation.

19. The method of claim 1 wherein step 1(c) comprises maintaining the sample droplet in a substantially stationary position for a period of time sufficient to permit coagulation.

20. The method of claim 1 wherein step 1(c) comprises heating the sample droplet.

21. The method of claim 1 wherein step 1(c) comprises cooling the sample droplet.

22. The method of claim 1 wherein step 1(c) is accomplished in the presence of an electrical field.

23. The method of claim 1 further comprising conducting an assay using the coagulated droplet as input.

24. The method of claim 1 wherein the method is effected using droplet operations on a droplet actuator.

25. The method of claim 1 wherein the method is effected using droplet operations in a droplet operations gap on a droplet actuator.

26. The method of claim 1 wherein the liquid filler fluid comprises a silicone oil, a carbon oil, and/or a fluorinated oil.

27. The method of claim 1 wherein the liquid filler fluid has a viscosity ranging from about 1 to about 3 cSt.

28. The method of claim 1 wherein the liquid filler fluid is doped with a surfactant.

29. The method of claim 28 wherein the surfactant comprises a linoleic acid based surfactant composition.

30-136. -136. (canceled)