Methods and systems for cell and bead processing

Patent metadata

Field	Value
Publication number	US-12117378-B2
Application number	US-202318205397-A
Country	US
Kind code	B2
Filing date	Jun 2, 2023
Priority date	Jun 25, 2018
Publication date	Oct 15, 2024
Grant date	Oct 15, 2024

Abstract

Methods and systems for cell and bead processing or analysis are provided. A cell or bead can be subjected to conditions sufficient to change a first characteristic or set of characteristics (e.g., cell or bead size). The cell or bead can further be subjected to conditions sufficient to change a second characteristic or set of characteristics. In some cases, crosslinks are formed within the cell or bead.

First claim

What is claimed is:

1. A method comprising:
(a) subjecting a first polymer matrix encasing a cell or a cell component to first conditions sufficient to reduce a first volume or a first diameter of said first polymer matrix, thereby yielding a first reduced polymer matrix encasing said cell or said cell component;
(b) subjecting a second polymer matrix to second conditions sufficient to reduce a second volume or a second diameter of said second polymer matrix, thereby yielding a second reduced polymer matrix; and
(c) co-partitioning said first reduced polymer matrix and said second reduced polymer matrix into a partition.

2. The method of claim 1 , wherein said method reduces said first diameter of said first polymer matrix.

3. The method of claim 1 , wherein said method reduces said first volume of said first polymer matrix.

4. The method of claim 1 , wherein said subjecting said first polymer matrix to said first conditions comprises bringing said first polymer matrix in contact with a chemical species, wherein said chemical species reduces said first volume or said first diameter of said first polymer matrix.

5. The method of claim 4 , wherein said chemical species comprises an organic solvent.

6. The method of claim 5 , wherein said organic solvent is acetone or an alcohol.

7. The method of claim 1 , wherein said subjecting said first polymer matrix to said first conditions comprises changing a temperature.

8. The method of claim 1 , wherein said first polymer matrix comprises poly (N-isopropylacrylamide).

9. The method of claim 1 , wherein said first polymer matrix comprises said cell component, and wherein said cell component comprises two or more cell components selected from a group consisting of a protein, a DNA, and an RNA.

10. The method of claim 9 , wherein said two or more cell components comprise a cross-link that couples said two or more cell components.

11. The method of claim 1 , wherein said method reduces said first volume or said first diameter by at least 6%.

12. The method of claim 11 , wherein said method reduces said first volume or said first diameter by at least 50%.

13. The method of claim 1 , wherein said subjecting said first polymer matrix to said first conditions further changes a fluidity characteristic or a rigidity characteristic of said first polymer matrix.

14. The method of claim 1 , wherein said subjecting said first polymer matrix to said first conditions further changes a porosity of said first polymer matrix.

15. The method of claim 1 , wherein said subjecting said first polymer matrix to said first conditions further changes a refractive index of said first polymer matrix.

16. The method of claim 1 , wherein said cell or said cell component is covalently attached to said first polymer matrix.

17. The method of claim 1 , wherein said cell or said cell component is noncovalently attached to said first polymer matrix.

18. The method of claim 1 , wherein said first polymer matrix comprises said cell.

19. The method of claim 18 , further comprising, during (a), lysing said cell within said first polymer matrix or said first reduced polymer matrix.

20. The method of claim 18 , further comprising lysing said cell within said partition.

21. The method of claim 1 , wherein said partition comprises a plurality of nucleic acid barcode molecules.

22. The method of claim 21 , wherein said first polymer matrix comprises said cell component, and wherein the method further comprises using a nucleic acid barcode molecule of said plurality of nucleic acid barcode molecules and said cell component to generate a barcoded cell component.

23. The method of claim 1 , wherein said second polymer matrix comprises a plurality of nucleic acid barcode molecules.

24. The method of claim 23 , wherein said first polymer matrix comprises said cell component, and wherein said method further comprises, after (c), generating a barcoded cell component using a nucleic acid barcode molecule of said plurality of nucleic acid barcode molecules and said cell component.

25. The method of claim 24 , wherein said generating said barcoded cell component occurs in said partition.

26. The method of claim 1 , wherein said first conditions and said second conditions are same conditions.

27. The method of claim 1 , wherein said first conditions and said second conditions are different conditions.

Assignees

• 10X Genomics Inc

Inventors

• Hindson Christopher
• Price Andrew D
• Schnall-Levin Michael

Classifications

• C12N5/0006

  Modification of the membrane of cells, e.g. cell decoration · CPC title

• G01N1/4077

  by other techniques involving separation of suspended solids · CPC title

• C12N2537/10

  Cross-linking · CPC title

• C12M35/08

  Chemical, biochemical or biological means, e.g. plasma jet, co-culture · CPC title

• C12M25/01

  Drops · CPC title