Droplet creation techniques
The present invention is generally related to systems and methods for producing droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, at least one droplet is used to create a plurality of droplets, using techniques such as flow-focusing techniques. In one set of embodiments, a plurality of droplets, containing varying species, can be divided to form a collection of droplets containing the various species therein. A collection of droplets, according to certain embodiments, may contain various subpopulations of droplets that all contain the same species therein. Such a collection of droplets may be used as a library in some cases, or may be used for other purposes.
1. A method for forming a plurality of droplets, comprising:
providing an emulsion comprising a plurality of droplets, at least some of the droplets comprising a first fluid the droplets substantially surrounded by a second fluid; and
passing the emulsion droplets through an intersection in a microfluidic channel to form a plurality of divided droplets, wherein the intersection comprises at least two intersecting channels containing an entering third fluid intersecting the microfluidic channel at the intersection.
2. The method of claim 1 , wherein the first fluid and the second fluid are substantially immiscible.
3. The method of claim 1 , wherein the plurality of divided droplets are substantially surrounded by the second fluid.
4. The method of claim 1 , wherein providing the emulsion comprises providing a plurality of droplets, each substantially surrounded by the second fluid, and passing the droplets comprises passing at least some of the droplets through the intersection of the microfluidic channel such that each of the droplets is divided to form two or more of the divided droplets.
5. The method of claim 1 , wherein providing the emulsion comprises providing a plurality of droplets, each substantially surrounded by the second fluid, and passing the droplets comprises passing at least some of the droplets through the intersection of the microfluidic channel such that each of the droplets is divided to form substantially the same number of divided droplets.
6. The method of claim 4 , wherein, for each of the droplets passing through the intersection of the microfluidic channel, the two or more divided droplets that are formed from each of the droplets have a distribution of diameters such that no more than about 5% of the divided droplets have a diameter greater than about 10% of the average diameter of all of the divided droplets that are formed.
7. The method of claim 5 , wherein the plurality of divided droplets has a distribution of diameters such that no more than about 5% of the droplets have a diameter greater than about 10% of the average diameter of the droplets.
8. The method of claim 5 , wherein the plurality of droplets contains therein at least four distinguishable species, such that no more than about 5% of the droplets contains two or more of the at least four distinguishable species therein.
9. The method of claim 8 , wherein the at least four distinguishable species comprises at least four distinguishable nucleic acids.
10. The method of claim 8 , wherein the at least four distinguishable species comprises at least four distinguishable identification elements.
11. The method of claim 8 , wherein the at least four distinguishable species comprises at least four distinguishable proteins.
12. The method of claim 1 , wherein the at least one droplet has an average diameter greater than about 500 microns and the plurality of divided droplets has an average diameter of less than about 500 microns.
13. The method of claim 1 , wherein at least about 10 divided droplets are formed from the at least one first droplet.
14. The method of claim 1 , wherein at least about 50 divided droplets are formed from the at least one first droplet.
15. The method of claim 1 , wherein the average diameter of the plurality of divided droplets is less than about 1000 microns and wherein the droplets are substantially monodisperse.
16. The method of claim 1 , wherein providing the emulsion comprises providing a first droplet comprising at least one first species and a second droplet comprising at least one second species distinguishable from the first species.
17. The method of claim 16 , further comprising forming an emulsion comprising the plurality of divided droplets, each divided droplet comprising a portion of the first droplet or a portion of the second droplet, wherein the average diameter of the divided droplets in the emulsion is less than about 1000 microns.
18. The method of claim 1 , wherein the third fluid is substantially identical to the second fluid.
19. A method for forming a plurality of droplets, comprising:
providing an emulsion comprising a plurality of droplets comprising a first fluid substantially surrounded by a second fluid; and
passing the emulsion through a microfluidic channel to form a plurality of divided droplets, wherein substantially each of the divided droplets contains therein at least four distinguishable identification elements.