Granted Patent B1

US 12,686,949 · App. 19/566,399 · Granted Jul 21, 2026

Inventors: Solongo B. Ziraldo (Pleasanton, CA); Geoffrey McDermott (Livermore, CA); Shea Thompson Lance (Livermore, CA)

Assignee: 10x Genomics, Inc.

C40B40/06

Abstract

Claims (19)

1 . A system for droplet processing, the system comprising:

a partitioning fluid comprising a fluorinated oil and a fluorosurfactant, wherein the fluorosurfactant comprises a perfluoropolyether (PFPE);

a first aqueous fluid comprising an ammonium salt and a surfactant, wherein the surfactant is sodium dodecyl sulfate (SDS); and

a plurality of beads, wherein a bead of the plurality of beads is coupled to a plurality of nucleic acid barcode molecules comprising at least 1,000 nucleic acid barcode molecules;

wherein the partitioning fluid and the first aqueous fluid are capable of forming a first emulsion comprising a first plurality of droplets and a plurality of micelles dispersed in the fluorinated oil, wherein the first plurality of droplets comprises a first droplet, the first droplet comprising the first aqueous fluid and a first droplet surface comprising the fluorosurfactant, and

wherein a micelle of the plurality of micelles is derived from the first droplet and comprises the surfactant;

wherein the partitioning fluid and a second aqueous fluid comprising the plurality of beads and a plurality of cells are capable of forming a second emulsion comprising a second plurality of droplets comprising a second droplet, the second droplet comprising the bead of the plurality of beads, a cell of the plurality of cells, and a second droplet surface comprising the fluorosurfactant; and

wherein the micelle is capable of transferring the surfactant to the second droplet after contacting the first emulsion with the second emulsion.

2 . The system of claim 1 , wherein the fluorosurfactant comprises a carboxylic acid-terminated PFPE.

3 . The system of claim 2 , wherein the fluorosurfactant is a first fluorosurfactant and the partitioning fluid further comprises a second fluorosurfactant.

4 . The system of claim 3 , wherein the second fluorosurfactant is a polyethylene glycol (PEG)-PFPE block copolymer.

5 . The system of claim 3 , wherein the second fluorosurfactant is a PFPE-polyethylene glycol (PEG)-PFPE triblock copolymer.

6 . The system of claim 3 , wherein the first fluorosurfactant or the second fluorosurfactant inhibits droplet coalescence.

7 . The system of claim 1 , wherein the first aqueous fluid further comprises n-dodecyl-beta-D-maltoside (DBDM), sodium lauroyl sarcosinate, t-Octylphenoxypolyethoxyethanol, or polysorbate 20.

8 . The system of claim 1 , wherein the partitioning fluid and the first aqueous fluid are capable of forming the first emulsion when combined and agitated.

9 . The system of claim 8 , wherein the partitioning fluid and the second aqueous fluid are capable of forming the second emulsion when combined and agitated.

10 . The system of claim 1 , wherein a nucleic acid barcode molecule of the plurality of nucleic acid barcode molecules comprises a bead-specific barcode sequence and a sequence capable of hybridizing to a messenger ribonucleic acid (mRNA) of the cell.

11 . The system of claim 10 , wherein the bead is a hydrogel bead.

12 . The system of claim 1 , wherein the micelle of the plurality of micelles further comprises the fluorosurfactant.

Continuity (5)

References Cited (400)