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

Patent metadata

Field	Value
Publication number	US-10150117-B2
Application number	US-201715717847-A
Country	US
Kind code	B2
Filing date	Sep 27, 2017
Priority date	Apr 10, 2014
Publication date	Dec 11, 2018
Grant date	Dec 11, 2018

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 device for generating a droplet comprising a single microcapsule of a plurality of microcapsules, said device comprising a layered structure comprising:
(i) a first channel connected to a first end of a flow regulator, wherein said first channel is configured to receive a first fluid comprising said plurality of microcapsules, wherein said microcapsules of said first fluid flow at an irregular frequency in said first channel;
(ii) a second channel connected to a droplet generation junction, wherein said second channel is configured to receive a second fluid that is immiscible with said first fluid;
(iii) a third channel connected to a droplet generation junction; and
(iv) a fourth channel connected to a second end of the flow regulator and to the droplet generation junction,
wherein said flow regulator comprises a broadened channel region at said first end and a funneling channel region ending at said second end, wherein said flow regulator is configured to (a) allow microcapsules of said plurality of microcapsules from said first channel to gather in said broadened channel region, and (b) allow said microcapsules gathered in said broadened channel region to flow through said funneling channel region and into said fourth channel at a regular flow frequency,
wherein said fourth channel is configured to bring said microcapsules in contact with said second fluid from said second channel at said droplet generation junction to generate a droplet comprising said single microcapsule from said microcapsules of said plurality of microcapsules, and
wherein said third channel is configured to provide an outlet for said droplet comprising said single microcapsule away from said droplet generation junction.

2. The device of claim 1 , wherein said first channel, said second channel, said third channel, and said fourth channel are in a first layer of said layered structure.

3. The device of claim 2 , wherein a first reservoir configured to contain said first fluid and a second reservoir configured to contain said second fluid are in a second layer of said layered structure, wherein said second layer is different than said first layer.

4. The device of claim 3 , wherein said second layer comprises a planar surface.

5. The device of claim 4 , wherein said planar surface of said second layer is mated with a planar surface of said first layer such that said first reservoir is connected to an end of said first channel and said second reservoir is connected to an end of said second channel.

6. The device of claim 5 , wherein said first reservoir has a tapering or conical shape towards said end of said first channel.

7. The device of claim 2 , wherein said first layer comprises a planar surface in which is disposed a series of channels or grooves that correspond to said first channel, said second channel, said third channel, and said fourth channel.

8. The device of claim 7 , further comprising a laminating structure adjacent to said first layer, wherein said laminating structure seals said series of channels or grooves.

9. The device of claim 1 , wherein said first channel, said second channel, said third channel, and said fourth channel are injection molded as a single part.

10. The device of claim 1 , wherein said flow regulator is dimensioned to provide said microcapsules in said fourth channel at a flow frequency is greater than 50 Hz at a coefficient of variation of less than 30%.

11. The device of claim 1 , wherein said flow regulator is dimensioned to provide said microcapsules in said fourth channel at a flow frequency that is sufficient to yield a plurality of droplets at said droplet generation junction, wherein said plurality of droplets has an occupancy of no more than 1 microcapsule per droplet.

12. The device of claim 1 , wherein said broadened channel region of said flow regulator narrows along a direction of flow and said funneling channel region rejoins a cross-sectional dimension of said fourth channel.

13. The device of claim 1 , wherein said flow regulator comprises a tapering or conical shape.

14. The device of claim 1 , wherein said layered structure further comprises a fifth channel, wherein said fifth channel is configured to receive a third fluid comprising an aqueous fluid.

15. The device of claim 14 , wherein (i) said first channel or said fourth channel and (ii) said fifth channel meet at a second junction upstream of said droplet generation junction.

16. The device of claim 14 , wherein said fifth channel comprises a passive fluid valve that is configured to break capillary forces of an aqueous fluid of said third fluid.

17. The device of claim 1 , wherein said first channel or said fourth channel comprises a passive fluid valve that is configured to break capillary forces of an aqueous fluid of said first fluid source.

18. A device for generating a droplet comprising a single microcapsule of a plurality of microcapsules, comprising
a layered structure comprising:
(i) a first channel configured to receive a first fluid comprising said plurality of microcapsules, wherein said first channel comprises a first region comprising a plurality of pillars that is configured to provide controlled dispensing of said plurality of microcapsules;
(ii) a second channel connected to a droplet generation junction, wherein said second channel is configured to receive a second fluid that is immiscible with said first fluid; and
(iii) a third channel connected to said droplet generation junction;
wherein said first channel is configured to bring microcapsules of said plurality of microcapsules in contact with said second fluid from said second channel at said droplet generation junction to generate a droplet comprising a single microcapsule from said microcapsules of said plurality of microcapsules, and
wherein said third channel is configured to provide an outlet for said droplet comprising said single microcapsule away from said droplet generation junction.

19. The device of claim 18 , wherein said first channel further comprises posts or tortuous paths.

20. The device of claim 18 , wherein said first channel, said second channel, and said third channel are in a first layer of said layered structure.

21. The device of claim 20 , wherein a first reservoir configured to contain said first fluid and a second reservoir configured to contain said second fluid are in a second layer of said layered structure, wherein said second layer is different than said first layer.

22. The device of claim 21 , wherein said second layer comprises a planar surface.

23. The device of claim 22 , wherein said planar surface of said second layer is mated with a planar surface of said first layer such that said first reservoir is connected to an end of said first channel and said second reservoir is connected to an end of said second channel.

24. The device of claim 20 , wherein said first layer comprises a planar surface in which is disposed a series of channels or grooves that correspond to said first channel, said second channel, and said third channel.

25. The device of claim 24 , further comprising a laminating structure adjacent to said first layer, wherein said laminating structure seals said series of channels or grooves.

26. The device of claim 18 , wherein said layered structure further comprises a fourth channel, wherein said fourth channel is configured to receive a third fluid comprising an

Assignees

• 10X Genomics Inc

Inventors

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

Classifications

• B01L2200/0673

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

• 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

• G01N2035/1034

  Transferring microquantities of liquid · CPC title

• B01L2400/086

  using baffles or other fixed flow obstructions · CPC title

• B01L3/0241 Primary

  Drop counters; Drop formers · CPC title