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

Patent metadata

Field	Value
Publication number	US-10137449-B2
Application number	US-201715717840-A
Country	US
Kind code	B2
Filing date	Sep 27, 2017
Priority date	Apr 10, 2014
Publication date	Nov 27, 2018
Grant date	Nov 27, 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 method for manufacturing a droplet generator, comprising:
(a) injection molding a polymeric 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 a 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;
(iii) a third channel connected to the droplet generation junction; and
(iv) a fourth channel connected to a second end of the flow regulator and to the droplet generation junction,
wherein a cross-section of said flow regulator (i) increases along a direction of flow from said first channel and decreases along a direction of flow to said fourth channel and (ii) is dimensioned to accept microcapsules of said plurality of microcapsules from said first channel and provide said microcapsules in said fourth channel;
wherein said fourth channel is configured to allow contact of a fluid comprising said microcapsules of said plurality of microcapsules 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 from said droplet generation junction; and

(b) attaching a laminating structure to said polymeric structure, wherein said laminating structure seals said first channel, said second channel, said third channel, and said fourth channel.

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

3. The method of claim 2 , wherein a first reservoir corresponding to said first fluid and a second reservoir corresponding to said second fluid are in a second layer of said polymeric structure, which second layer is different than said first layer.

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

5. The method of claim 4 , wherein said first reservoir has a tapering or conical shape towards said end of said first channel.

6. The method 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.

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

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

9. The method 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 a an occupancy of no more than 1 microcapsule per droplet.

10. The method of claim 1 , wherein said flow regulator comprises a funneling structure.

11. The method of claim 1 , wherein said flow regulator includes a broadened region that narrows along a direction of flow from said first channel to said fourth channel.

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

13. The method of claim 1 , wherein said polymeric structure further comprises a fifth channel, wherein said fifth channel is configured to receive a third fluid comprising an aqueous fluid.

14. The method of claim 13 , wherein said (i) first channel or said fourth channel and (ii) said fifth channel meet at a second junction upstream of said droplet generation junction.

15. The method of claim 13 , wherein said fifth channel comprises a passive fluid valve that is configured to break capillary forces of an aqueous fluid of said third fluid.

16. The method 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.

17. The method of claim 1 , wherein said first channel and said fourth channel are part of the same channel.

18. The method of claim 1 , wherein at least a portion of said cross-section of said flow regulator gradually decreases along a direction of flow to said fourth channel.

19. A method for manufacturing a droplet generator, comprising:
(a) injection molding a polymeric structure comprising:
(i) a first channel configured to receive a first fluid comprising a 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 the droplet generation junction,
wherein said first channel is configured to allow contact of a fluid comprising said microcapsules of said plurality of microcapsules 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 from said droplet generation junction; and

(b) attaching a laminating structure to said polymeric structure, wherein said laminating structure seals said first channel, said second channel, and said third channel.

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

21. The method of claim 19 , wherein said first channel, said second channel and said third channel are in a first layer of said polymeric structure.

22. The method of claim 21 , wherein a first reservoir corresponding to said first fluid and a second reservoir corresponding to said second fluid are in a second layer of said polymeric structure, which second layer is different than said first layer.

23. The method of claim 22 , wherein said second layer comprises a planar surface, said first reservoir, and said second reservoir.

24. The method of claim 23 , 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.

25. The method of claim 21 , 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.

26. The method of claim 15 , wherein said polymeric structure further comprises a fourth channel, wherein said fourth channel is configured to receive a third flui

Assignees

• 10X Genomics Inc

Inventors

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

Classifications

• B01L2400/086

  using baffles or other fixed flow obstructions · CPC title

• B01L2400/049

  vacuum · 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

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title

• G01N2035/00237

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