Apparatus and methods for multi-step channel emulsification

Patent metadata

Field	Value
Publication number	US-10662470-B2
Application number	US-201615070108-A
Country	US
Kind code	B2
Filing date	Mar 15, 2016
Priority date	Mar 16, 2015
Publication date	May 26, 2020
Grant date	May 26, 2020

Abstract

Methods and devices for forming droplets are provided. In certain embodiments, the methods and devices form droplets having different diameters.

First claim

We claim:

1. An emulsification device comprising:
a channel having an inlet portion having a channel height CH and a width CW, wherein the ratio of CW/CH is greater than 0.2 and less than 5.0;
a first step in fluid communication with the inlet portion, wherein:
the first step has a tread length T 1 and a step height SH 1 ;
SH 1 is greater than CH by a riser height R 1 ; and
the ratio of SH 1 /CH is greater than 1.0 and less than 5.0;

a second step in fluid communication with the first step, wherein:
the second step has a tread length T 2 and a step height SH 2 ;
SH 2 is greater than SH 1 by a riser height R 2 ;
the ratio of SH 2 /CH is greater than 1.0 and less than 5.0; and
the ratio of T 2 /CH is less than the ratio of T 1 /CH;

a third step in fluid communication with the second step, wherein:
the third step has a step height SH 3 ;
SH 3 is greater than SH 2 by a riser height R 3 ; and
R 3 is greater than zero.

2. The emulsification device of claim 1 wherein the ratio of SH 1 /CH is greater than 1.0 and less than 2.0.

3. The emulsification device of claim 1 wherein R 3 is greater than 50 microns.

4. The emulsification device of claim 1 wherein the ratio of T 1 /CH is between 3.0 and 4.0.

5. The emulsification device of claim 1 wherein the ratio of T 2 /CH is between 2.0 and 4.0.

6. The emulsification device of claim 1 wherein CH is between 10 microns and 50 microns.

7. The emulsification device of claim 1 further comprising:
a plurality of inlet portions, wherein each inlet portion in the first plurality of inlet portions has a height CH and a width CW, and wherein the ratio of CW/CH is greater than 0.2 and less than 5.0.

8. The emulsification device of claim 7 wherein the plurality of inlet portions comprises between 10 and 100 inlet portions.

9. A method of forming an emulsion, the method comprising:
obtaining an emulsification device according to claim 7 , wherein the plurality of first steps, the plurality of second steps and the plurality of third steps contain a first fluid that is substantially static; and
introducing a second fluid into the plurality of inlet portions and through the plurality of first steps, the plurality of second steps and the third step, wherein:
a partial droplet of the second fluid forms in each of the plurality of first steps;
a complete droplet of the second fluid forms during the transition between the plurality of first steps and the second steps in each of the plurality of second steps; and
the complete droplet of the second fluid is directed from the plurality of second steps to the third step.

10. The method of claim 9 , wherein at least 10,000 complete droplets are directed from the plurality of second steps to the third step per minute.

11. The method of claim 10 , wherein the droplets have an average dispersion of less than 10 percent.

12. The method of claim 9 , wherein the average droplet diameter of droplets in the third step is between 40 to 300 microns.

13. The emulsification device of claim 7 further comprising:
a plurality of first steps, wherein each first step in the plurality of first steps is:
in fluid communication with each of the plurality of inlet portions; and
has a length T 1 and a height SH 1 , wherein SH 1 is greater than CH by a riser height R 1 , and wherein the ratio of SH 1 /CH is greater than 1.0 and less than 5.0.

14. The emulsification device of claim 13 further comprising:
a plurality of second steps, wherein each second step in the plurality of second steps is:
in fluid communication with a first step in the plurality of first steps;
in fluid communication with the third step; and
has a length T 2 and a height SH 2 , wherein SH 2 is greater than SH 1 by a riser height R 2 , and wherein the ratio of SH 2 /CH is greater than 1.0 and less than 5.0.

15. The emulsification device of claim 7 further comprising:
a single continuous first step, wherein the single continuous first step is:
in fluid communication with each of the plurality of inlet portions; and
has a length T 1 and a height SH 1 , wherein SH 1 is greater than CH by a riser height R 1 , and wherein the ratio of SH 1 /CH is greater than 1.0 and less than 5.0.

16. The emulsification device of claim 15 further comprising:
a single continuous second step, wherein the single continuous second step is:
in fluid communication with the single continuous first step;
in fluid communication with the third step; and
has a length T 2 and a height SH 2 , wherein SH 2 is greater than SH 1 by a riser height R 2 , and wherein the ratio of SH 2 /CH is greater than 1.0 and less than 5.0.

17. A method of forming an emulsion, the method comprising:
obtaining an emulsification device according to claim 1 , wherein the first step, the second step and the third step contain a first fluid that is substantially static; and
introducing a second fluid into the inlet portion and through the first step, the second step and the third step, wherein:
a partial droplet of the second fluid forms in the first step;
a complete droplet of the second fluid forms in the second step; and
the complete droplet of the second fluid is directed from the second step to the third step.

18. The method of claim 17 wherein the second fluid contains an analyte of interest and an assay reagent.

19. The method of claim 17 wherein the first fluid is a hydrophobic liquid and the second fluid is a hydrophilic liquid.

20. The method of claim 17 wherein the first fluid is a hydrophilic liquid and the second fluid is a hydrophobic liquid.

21. The method of claim 17 wherein the either the first fluid or the second fluid comprises an emulsifying agent.

22. The method of claim 21 wherein the emulsifying agent comprises a non-ionic surfactant or a blocking protein.

23. The method of claim 17 wherein a complete droplet of the second fluid forms in the second step at a rate of between 1 and 30 complete droplets per second.

24. The method of claim 17 wherein the complete droplet of second fluid has an average diameter between 40 and 300 microns.

25. The method of claim 17 wherein the emulsion formed between the first fluid and the second fluid has a monodispersity between two and ten percent.

Assignees

• Luminex Corp

Inventors

• Arab Nicolas
• Estrada Arnold
• Fine Daniel
• Johnson Ross

Classifications

• B01L7/52

  with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples · CPC title

• B01L3/502784

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

• C12Q1/686 Primary

  Polymerase chain reaction [PCR] · CPC title

• B01F3/0807

  Operations & Transport · mapped topic

• B01F3/0865

  Operations & Transport · mapped topic