Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same

Patent metadata

Field	Value
Publication number	US-10343166-B2
Application number	US-201715596754-A
Country	US
Kind code	B2
Filing date	May 16, 2017
Priority date	Apr 10, 2014
Publication date	Jul 9, 2019
Grant date	Jul 9, 2019

Abstract

The disclosure provides devices, systems and methods for the generation of encapsulated reagents and the partitioning of encapsulated reagents for use in subsequent analyses and/or processing, such as in the field of biological analyses and characterization.

First claim

What is claimed is:

1. A method for partitioning microcapsules, comprising:
(a) providing a partitioning system comprising:
(i) a first channel comprising a plurality of filtering structures that is configured to receive a first fluid comprising a plurality of microcapsules;
(ii) a second channel configured to receive a second fluid that is immiscible with the first fluid;
(iii) a third channel; and
(iv) a droplet generation junction connected to the first channel, the second channel, and the third channel; and

(b) in the partitioning system,
(i) subjecting the first fluid to flow along the first channel and through the plurality of filtering structures such that the first fluid is filtered and microcapsules of the plurality of microcapsules flow along the first channel to the droplet generation junction, and
(ii) subjecting the second fluid to flow along the second channel towards the droplet generation junction, wherein the first fluid and the second fluid meet at the droplet generation junction to generate a plurality of droplets comprising the microcapsules of the plurality of microcapsules, which plurality of droplets flow along the third channel.

2. The method of claim 1 , wherein the microcapsules of the plurality of microcapsules flow along the first channel at aUthell flow frequency that has a coefficient of variation of less than 30%.

3. The method of claim 2 , wherein the flow frequency has a coefficient of variation of less than 10%.

4. The method of claim 3 , wherein the flow frequency has a coefficient of variation of less than 5%.

5. The method of claim 1 , wherein the microcapsules of the plurality of microcapsules flow along the first channel at affthell flow frequency that is greater than 50 Hz.

6. The method of claim 5 , wherein the flow frequency is greater than 1000 Hz.

7. The method of claim 6 , wherein the flow frequency is greater than 2000 Hz.

8. The method of claim 1 , wherein the first channel, second channel, and third channel are configured to provide the microcapsules of the plurality of microcapsules in the plurality of droplets at an occupancy of no more than 1 microcapsule per droplet.

9. The method of claim 1 , wherein the microcapsules of the plurality of microcapsules flow along the first channel at differenehell flow frequencies upstream and downstream of the plurality of filtering structures.

10. The method of claim 1 , wherein first fluid is an aqueous fluid.

11. The method of claim 1 , wherein the plurality of microcapsules comprises one or more reagents.

12. The method of claim 11 , wherein the one or more reagents are encapsulated in the plurality of microcapsules.

13. The method of claim 11 , wherein the one or more reagents are barcode sequences.

14. The method of claim 1 , wherein the plurality of microcapsules is monodisperse.

15. The method of claim 14 , wherein microcapsules of the plurality of microcapsules have a mean cross-sectional dimension and a coefficient of variation in cross-sectional dimension of no greater than 30%.

16. The method of claim 1 , wherein the first fluid and the second fluid are subjected to flow into the droplet generation junction at flow rates that generate the plurality of droplets such that a given droplet of the plurality of droplets comprises a single microcapsule of the plurality of microcapsules.

17. The method of claim 1 , wherein the partitioning system further comprises a fourth channel that is configured to receive a third fluid comprising an additional aqueous fluid.

18. The method of claim 17 , wherein the first channel and the fourth channel meet at a junction upstream of the droplet generation junction, and the third fluid flows along the fourth channel to the junction, thereby bringing the first fluid and the third fluid in contact to form a mixture comprising the first fluid and the additional aqueous fluid.

19. The method of claim 18 , wherein the junction and the droplet generation junction are connected through a fifth channel.

20. The method of claim 1 , wherein microcapsules of the plurality of microcapsules are solid particles.

21. The method of claim 1 , wherein microcapsules of the plurality of microcapsules are gel particles.

22. The method of claim 1 , wherein the first fluid and the second fluid are subjected to flow substantially simultaneously.

23. The method of claim 1 , wherein the plurality of filtering structures comprise a plurality of pillars, posts, or torturous paths.

24. The method of claim 1 , wherein the plurality of filtering structures are disposed in a region of the first channel, which region is disposed at an end of the first channel opposite an end connected to the droplet generation junction.

25. The method of claim 1 , wherein the plurality of filtering structures provide controlled dispensing of the microcapsules of the plurality of microcapsules in the first channel.

26. The method of claim 10 , wherein the aqueous fluid comprises a biological molecule.

Assignees

• 10X Genomics Inc

Inventors

• Bharadwaj Rajiv
• Ness Kevin
• Mitra Debkishore
• Masquelier Donald
• Makarewicz Anthony
• Hindson Christopher
• Hindson Benjamin
• Saxonov Serge

Classifications

• B01L3/502761 Primary

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• B01L3/502784

  specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title

• B01L2200/0673

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

• G01N35/085

  Flow Injection Analysis · CPC title

• G01N2035/00237

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