Apparatus and methods for conducting assays and high throughput screening

Patent metadata

Field	Value
Publication number	US-10509018-B2
Application number	US-201514928631-A
Country	US
Kind code	B2
Filing date	Oct 30, 2015
Priority date	Nov 16, 2000
Publication date	Dec 17, 2019
Grant date	Dec 17, 2019

Abstract

The present invention provides microfluidic devices and methods for using the same. In particular, microfluidic devices of the present invention are useful in conducting a variety of assays and high throughput screening. Microfluidic devices of the present invention include elastomeric components and comprise a main flow channel; a plurality of branch flow channels; a plurality of control channels; and a plurality of valves. Preferably, each of the valves comprises one of the control channels and an elastomeric segment that is deflectable into or retractable from the main or branch flow channel upon which the valve operates in response to an actuation force applied to the control channel.

First claim

What is claimed is:

1. A method comprising:
a) providing a microfluidic device comprising flow channels;
b) providing a sample, wherein the sample comprises cells;
c) introducing the sample into a plurality of said flow channels of the microfluidic device so as to store the cells in a plurality of holding areas, such that there are a plurality of said holding areas in which a single cell is retained wherein each holding area is a segment of a flow channel formed by actuation of at least two valves;
d) detecting said single cells in the microfluidic device; and
e) treating said single cells with a detergent.

2. The method of claim 1 , further comprising assaying a property of the single cells.

3. The method of claim 2 , wherein the property is cell membrane permeability.

4. The method of claim 2 , wherein assaying a property of the single cells comprises applying a histological stain to cells of the sample.

5. The method of claim 2 , wherein assaying a property of the single cells comprises detecting protein markers expressed by the cells, wherein the protein markers are bound by labeled antibodies.

6. The method of claim 2 , wherein assaying a property of the single cells comprises mixing different test compounds with the cells.

7. The method of claim 6 , wherein assaying a property of the single cells further comprises detecting a reporter expression of the cells.

8. The method of claim 1 , wherein the assay mixture further comprises agents necessary to conduct RT-PCR.

9. The method of claim 1 , further comprising performing an RT-PCR reaction.

10. The method of claim 8 , further comprising performing an RT-PCR reaction.

11. The method of claim 1 , wherein in step c) the cells are introduced using a multichannel robotic pipettor.

12. The method of claim 1 , wherein step d) of detecting is performed by an optical detector that is a separate element from the microfluidic device.

13. The method of claim 12 , wherein the optical detector comprises one or more photodiodes.

14. The method of claim 12 , wherein the optical detector comprises an optical microscope.

15. The method of claim 12 , wherein the optical detector comprises a CCD camera.

16. The method of claim 1 , wherein step d) of detecting comprises scanning the microfluidic device under a microscope objective, wherein the microfluidic device is attached to a translatable stage.

17. The method of claim 1 , wherein step d) of detecting comprises detecting at least one of fluorescence intensity, fluorescence polarization, fluorescence resonance energy transfer, and fluorescence correlation.

18. The method of claim 2 , wherein assaying a property of the single cells comprises detecting at least one of fluorescence intensity, fluorescence polarization, fluorescence resonance energy transfer, and fluorescence correlation.

19. The method of claim 1 , wherein the microfluidic device comprises an elastomeric material characterized by a Young's modulus between 1 Pa and 1 TPa.

20. The method of claim 1 wherein the single cells are treated with an assay mixture that comprises the detergent along with primers and a polymerase, wherein said polymerase is selected from DNA polymerase and RNA polymerase.

21. The method of claim 20 , wherein said assay mixture further comprises an RNase inhibitor.

22. The method of claim 1 , wherein said single cells are detected in an enrichment section of the microfluidic device.

23. The method of claim 22 , wherein said enrichment section is a section of a flow channel.

24. The method of claim 1 , wherein one or more microfluidic systems within the microfluidic device comprises an elastomeric polymer.

25. The method of claim 1 wherein the valves comprise elastomeric segments that can be deflected into or out from a flow channel.

26. The method of claim 1 wherein the valves are actuated electrostatically or magnetically.

Assignees

• California Inst Of Techn

Inventors

• Quake Stephen R
• Unger Marc A
• Chou Hou-Pu
• Thorsen Todd A
• Scherer Axel

Classifications

• B01L3/502746

  characterised by the means for controlling flow resistance, e.g. flow controllers, baffles or throttle valves · CPC title

• G01N33/56966

  Animal cells · CPC title

• F16K99/0059

  actuated by a pilot fluid · CPC title

• B01L2300/1827

  using resistive heater · CPC title

• B01L2300/0877

  Flow chambers · CPC title