Method of producing emulsions

Patent metadata

Field	Value
Publication number	US-10583440-B2
Application number	US-201916443629-A
Country	US
Kind code	B2
Filing date	Jun 17, 2019
Priority date	Aug 22, 2017
Publication date	Mar 10, 2020
Grant date	Mar 10, 2020

Abstract

Devices, systems, and their methods of use, for generating droplets are provided. One or more geometric parameters of a microfluidic channel can be selected to generate droplets of a desired and predictable droplet size.

First claim

What is claimed is:

1. A method of producing droplets comprising:
a) providing a device comprising:
i) a first channel having a first proximal end, a first distal end, a first depth, and a first width, the first channel comprising a first liquid;
ii) a second channel having a second proximal end, a second distal end, a second depth, and a second width, wherein the second channel intersects the first channel between the first proximal end and the first distal end, the second channel comprising a third liquid;
iii) a shelf region comprising a second liquid and being in fluid communication with the first distal end, wherein the shelf region has a width greater than the first width; and
iv) a step region having a depth that is greater than the first depth and greater than the depth of the shelf region, wherein the shelf region is disposed between the first channel and the step region;
wherein the first liquid is immiscible with the second liquid, wherein the first or third liquid comprises one or more particles, and wherein the shelf region and the step region comprise a second liquid; and

b) combining the first liquid from the first channel and the third liquid from the second channel at the intersection; and
c) flowing the combined first and third liquids to the step region to produce droplets of the first liquid, the third liquid, and one or more particles, wherein the droplets are dispersed in the second liquid.

2. The method of claim 1 , wherein a proximal-to-distal direction extends from the first proximal end to the first distal end, and the first depth decreases in the proximal-to-distal direction in at least a portion of the first channel.

3. The method of claim 1 , wherein a proximal-to-distal direction extends from the first proximal end to the first distal end, and the first depth increases in the proximal-to-distal direction in at least a portion of the first channel.

4. The method of claim 1 , wherein the device further comprises a first reservoir in fluid communication with the first proximal end.

5. The method of claim 4 , wherein, prior to step a), the first liquid comprises the one or more particles, and the first reservoir comprises at least a portion of the first liquid and the one or more particles.

6. The method of claim 1 , wherein step c) produces droplets having a single particle.

7. The method of claim 1 , wherein the first liquid is aqueous or miscible with water.

8. The method of claim 1 , wherein the density of the first liquid is lower than the density of the second liquid.

9. The method of claim 1 , wherein the density of the first liquid is higher than the density of the second liquid.

10. The method of claim 1 , wherein a proximal-to-distal direction extends from the second proximal end to the second distal end, and the second depth decreases in the proximal-to-distal direction in at least a portion of the second channel.

11. The method of claim 1 , wherein a proximal-to-distal direction extends from the second proximal end to the second distal end, and the second depth increases in the proximal-to-distal direction in at least a portion of the second channel.

12. The method of claim 1 , wherein the third liquid is aqueous or miscible with water.

13. The method of claim 1 , wherein the density of the third liquid is lower than the density of the second liquid.

14. The method of claim 1 , wherein the density of the third liquid is higher than the density of the second liquid.

15. The method of claim 1 , wherein the device further comprises a second reservoir in fluid communication with the second proximal end.

16. The method of claim 1 , wherein a proximal-to-distal direction extends from the first distal end to the step region, and the shelf region has a width that increases in the proximal-to-distal direction.

17. The method of claim 1 , wherein the depth of the step region is at least 100 μm.

18. The method of claim 1 , wherein the droplets have a volume less than 1000 pL.

19. The method of claim 1 , wherein the one or more particles are beads.

20. The method of claim 19 , wherein, in step a), the first or third liquid comprises one or more biological particles, and the droplets produced in step c) comprise the first liquid, third liquid, a single bead, and a single biological particle.

Assignees

• 10X Genomics Inc

Inventors

• Bharadwaj Rajiv
• Makarewicz Anthony
• Lin Bill Kengli

Classifications

• B01L3/0241

  Drop counters; Drop formers · CPC title

• B01L2400/086

  using baffles or other fixed flow obstructions · CPC title

• C40B40/06

  Libraries containing nucleotides or polynucleotides, or derivatives thereof · CPC title

• G01N33/58

  involving labelled substances (G01N33/53 takes precedence) · CPC title

• B01L2200/0652

  Sorting or classification of particles or molecules · CPC title