Microfluidic devices

Patent metadata

Field	Value
Publication number	US-10639597-B2
Application number	US-201815886212-A
Country	US
Kind code	B2
Filing date	Feb 1, 2018
Priority date	May 11, 2006
Publication date	May 5, 2020
Grant date	May 5, 2020

Abstract

The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.

First claim

What is claimed is:

1. A method of analyzing a compound released from a cell, the method comprising:
providing a microfluidic substrate comprising an inlet module comprising a first inlet channel comprising a plurality of cells in aqueous solution and a second inlet channel comprising a plurality of primers in aqueous solution;
flowing the aqueous solutions together to form a combined aqueous solution, and flowing the combined aqueous solution through a nozzle into a reservoir comprising an oil, thereby extruding a plurality of droplets of aqueous solution into the reservoir, each droplet comprising primers and at least one cell from the plurality of cells;
incubating the plurality of the droplets off of the microfluidic substrate;
causing at least one secreted substance to be released from the cell in each droplet; and
conducting a homogenous assay with the primers on the secreted substance, the homogenous assay being conducted inside each of the plurality of droplets.

2. The method of claim 1 , wherein the cell is a bacterium cell, a fungal cell, a plant cell, or an animal cell.

3. The method of claim 1 , wherein each of the plurality of droplets in the providing step comprises a substrate for an enzyme.

4. The method of claim 3 , wherein the enzyme is conjugated to an antibody.

5. The method of claim 1 , wherein each of the plurality of droplets in the providing step further comprises a labeled compound.

6. The method of claim 1 , wherein the primers are bound to a bead.

7. The method of claim 1 , wherein the primers are barcoded.

8. The method of claim 1 , wherein each of the plurality of droplets in the providing step comprises reagents for an amplification reaction.

9. The method of claim 1 , wherein the secreted substance comprises a nucleic acid.

10. The method of claim 1 , wherein the homogenous assay comprises amplification.

11. The method of claim 10 , wherein amplification comprises PCR, QPCR, or rolling circle amplification.

12. The method of claim 5 , wherein the labeled compound comprises a dye.

13. The method of claim 1 , further comprising lysing the cell in each droplet before the conducting step.

14. The method of claim 1 , wherein the oil comprises a surfactant.

15. The method of claim 1 , further comprising breaking the droplets to release a product of the homogenous assay.

16. The method of claim 15 , further comprising sequencing the product of the homogenous assay.

Assignees

• Bio Rad Laboratories

Inventors

• Link Darren Roy
• Weiner Michael
• Marran David
• Rothberg Jonathan M

Classifications

• B03C5/026

  using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC] · CPC title

• G01N2035/00237

  Handling microquantities of analyte, e.g. microvalves, capillary networks · CPC title

• Y10T137/87652

  With means to promote mixing or combining of plural fluids · CPC title

• C12Q1/00

  Measuring or testing processes involving enzymes, nucleic acids or microorganisms (measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34); Compositions therefor; Processes of preparing such compositions · CPC title

• B01L2200/0673

  Handling of plugs of fluid surrounded by immiscible fluid · CPC title