Method of producing emulsions

Patent metadata

Field	Value
Publication number	US-10898900-B2
Application number	US-202016812866-A
Country	US
Kind code	B2
Filing date	Mar 9, 2020
Priority date	Aug 22, 2017
Publication date	Jan 26, 2021
Grant date	Jan 26, 2021

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 system comprising:
(A) a device comprising:
i) a plurality of first channels, each of the plurality of first channels having a first proximal end, a first distal end, a first depth, and a first width;
ii) a plurality of second channels, each of the plurality of second channels having a second proximal end, a second distal end, a second depth, and a second width, wherein each of the plurality of second channels intersects a corresponding one of the plurality of first channels between the first proximal end and the first distal end thereof; and
iii) a plurality of shelf regions and a plurality of step regions, each of the plurality of shelf regions having a shelf region depth, and each of the plurality of step regions having a step region depth, wherein the first distal end of at least one of the plurality of first channels is in fluid communication with one of the plurality of step regions via a corresponding one of the plurality of shelf regions, wherein each of the plurality of shelf regions has a shelf region width that is greater than the first width of the at least one of the plurality of first channels in fluid communication therewith, wherein each of the step region depths of the plurality of step regions is greater than the shelf region depth of the corresponding one of the plurality of shelf regions and the first depth of the at least one of the plurality of first channels in fluid communication therewith;

(B) a first liquid disposed in at least one of the plurality of first channels;
(C) a second liquid disposed in at least one of the plurality of shelf regions that is in fluid communication with the at least one of the plurality of first channels in (B); and
(D) a third liquid disposed in the at least one of the plurality of second channels that intersects the at least one of the plurality of first channels in (B);
wherein the first liquid is immiscible with the second liquid, and wherein the first or third liquid comprises one or more particles;
b) combining the first liquid of (B) and the third liquid of (D) at the intersection of the at least one of the plurality of first channels of (B) and the at least one of the plurality of second channels of (C) to provide a combined liquid; and
c) producing droplets of the combined liquid and at least one of the one or more particles by flowing the combined liquid to the one of the plurality of step regions that is in fluid communication with the at least one of the plurality of first channels of (B), 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 of each of the plurality of first channels, and the first depths of the plurality of first channels decrease in the proximal-to-distal direction in at least a portion of each of the plurality of first channels.

3. The method of claim 1 , wherein a proximal-to-distal direction extends from the first proximal end to the first distal end of each of the plurality of first channels, and the first depths of the plurality of first channels increase in the proximal-to-distal direction in at least a portion of each of the plurality of first channels.

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

5. The method of claim 4 , wherein the first reservoir further comprises at least a portion of 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 of each of the plurality of second channels, and the second depths of the plurality of second channels decrease in the proximal-to-distal direction in at least a portion of each of the plurality of second channels.

11. The method of claim 1 , wherein a proximal-to-distal direction extends from the second proximal end to the second distal end of each of the plurality of second channels, and the second depths of the plurality of second channels increase in the proximal-to-distal direction in at least a portion of each of the plurality of second channels.

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 of at least one of the plurality of second channels.

16. The method of claim 1 , wherein a proximal-to-distal direction extends from the first distal ends of each of the plurality of first channels to the corresponding step regions, and the shelf region widths increase in the proximal-to-distal direction.

17. The method of claim 1 , wherein each of the step region depths 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 combined liquid, a single bead, and a single biological particle.

Assignees

• 10X Genomics Inc

Inventors

• Bharadwaj Rajiv
• Makarewicz Anthony
• Lin Bill Kengli

Classifications

• B01F25/31422

  with a plurality of perforations in the axial direction only · CPC title

• B01F33/3021

  the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets · CPC title

• B01F33/3011

  using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream · CPC title

• B01F31/84

  for material continuously moving through a tube, e.g. by deforming the tube · CPC title

• B01F23/4105

  Methods of emulsifying · CPC title