Granted Patent B2

US 12,576,400 · App. 19/318,315 · Granted Mar 17, 2026

Inventors: Tarun Kumar Khurana (Fremont, CA); Ali Agah (Redwood City, CA); Yir-Shyuan Wu (Albany, CA); Filiz Gorpe Yasar (Redwood City, CA); Vernon A. Norviel (Evergreen, CO)

Assignee: Cellanome, Inc.

B01L3/502715B01L3/502707B01L3/502761C12Q1/6869C12Q1/6874G01N33/54386G01N33/6842B01L2200/0647B01L2200/12B01L2300/0663B01L2300/0883B01L2300/12B01L2300/163B01L2300/18C12Q2600/158G01N2570/00

Abstract

Claims (48)

1 . A method for analyzing a cell, comprising:

a) providing energy to a fluidic device comprising a fluid to polymerize a polymer precursor in the fluidic device, thereby forming a compartment comprising a polymer matrix wall within the fluid, wherein the fluid comprises a plurality of cells, wherein the compartment encloses a cell of the plurality of cells, and wherein the compartment physically separates the cell from an additional cell of the plurality of cells;

b) incubating the cell for at least about 1 minute, wherein the cell is inside of the compartment during the incubating; and

c) performing an assay within the compartment, thereby analyzing the cell, wherein the assay comprises:

lysing the cell, thereby releasing a nucleic acid from the cell;

capturing the nucleic acid using a capture element, wherein the capture element is inside of the compartment; and

reverse transcribing the nucleic acid to form a cDNA molecule.

2 . The method of claim 1 , wherein the compartment comprises a volume of about 100 picoliters to about 10 nanoliters.

3 . The method of claim 1 , wherein the assay comprises detecting the nucleic acid from the cell.

4 . The method of claim 1 , wherein the capture element comprises an oligonucleotide, a unique molecular identifier, a barcode, and a sequencing primer.

5 . The method of claim 1 , wherein the nucleic acid comprises a messenger ribonucleic acid (mRNA) molecule.

6 . The method of claim 1 , further comprising forming a plurality of additional compartments, wherein at least a subset of the plurality of additional compartments comprise additional cells of the plurality of cells.

7 . The method of claim 1 , wherein the fluidic device comprises a microfluidic device or a nanofluidic device.

8 . The method of claim 1 , wherein the method further comprises detecting a protein secreted by the cell.

9 . The method of claim 8 , wherein the detecting the protein secreted by the cell comprises:

capturing the protein on a capture antibody, wherein the capture antibody is inside of the compartment;

coupling a detection antibody to the protein; and

detecting the detection antibody.

10 . The method of claim 9 , wherein the capture antibody is coupled to a bead, and wherein the bead is inside of the compartment.

11 . The method of claim 1 , wherein the compartment encloses a second cell of the plurality of cells, and wherein the assay comprises detecting an interaction between the cell and the second cell.

12 . The method of claim 1 , wherein the polymer matrix wall extends from a first surface of the fluidic device to a second surface of the fluidic device opposite the first surface, thereby forming an interior of the compartment, wherein the interior of the compartment comprises the cell.

13 . The method of claim 1 , wherein the energy comprises light, and wherein the light is projected into the fluid using a spatial light modulator.

14 . The method of claim 13 , wherein the spatial light modulator comprises a digital micromirror device.

15 . The method of claim 13 , wherein during a), the light is projected to a location of the cell or adjacent to the location of the cell.

16 . The method of claim 1 , wherein the method further comprises imaging the cell.

17 . The method of claim 16 , wherein the imaging comprises bright-field imaging, phase-contrast imaging, fluorescence imaging, or a combination thereof.

18 . The method of claim 1 , wherein the cell is enclosed within the compartment with antibodies specific for surface proteins, and wherein the method further comprises determining a relative expression of the surface proteins on the cell.

19 . The method of claim 1 , wherein the plurality of cells are stationary during a).

20 . The method of claim 1 , wherein at least a subset of the plurality of cells are alive during a) and b).

21 . The method of claim 1 , wherein the plurality of cells are from a human subject, and wherein the method further comprises assessing a health status of the human subject at least in part based on the assay performed in (c).

22 . The method of claim 1 , wherein a) comprises crosslinking the polymer precursor to form the polymer matrix wall.

23 . A method for analyzing a cell, comprising:

a) providing energy to a fluidic device comprising a fluid to polymerize a polymer precursor in the fluidic device, thereby forming a compartment comprising a polymer matrix wall within the fluid, wherein the fluid comprises a plurality of cells, wherein the compartment encloses a cell of the plurality of cells, and wherein the compartment physically separates the cell from an additional cell of the plurality of cells;

b) incubating the cell for at least about 1 minute, wherein the cell is inside of the compartment during the incubating; and

c) performing an assay within the compartment, thereby analyzing the cell, wherein the assay comprises detecting a protein secreted by the cell.

24 . The method of claim 23 , wherein the detecting the protein secreted by the cell comprises:

capturing the protein on a capture antibody, wherein the capture antibody is inside of the compartment;

coupling a detection antibody to the protein; and

detecting the detection antibody.

25 . The method of claim 24 , wherein the capture antibody is coupled to a bead, and wherein the bead is inside of the compartment.

26 . A method for analyzing a cell, comprising:

a) providing energy to a fluidic device comprising a fluid to polymerize a polymer precursor in the fluidic device, thereby forming a compartment comprising a polymer matrix wall within the fluid, wherein the fluid comprises a plurality of cells, wherein the compartment encloses a cell of the plurality of cells with antibodies specific for surface proteins, and wherein the compartment physically separates the cell from an additional cell of the plurality of cells;

b) incubating the cell for at least about 1 minute, wherein the cell is inside of the compartment during the incubating; and

c) performing an assay within the compartment, thereby analyzing the cell, wherein the assay comprises determining a relative expression of the surface proteins on the cell.

27 . A method for analyzing a cell, comprising:

a) providing energy to a fluidic device comprising a fluid to polymerize a polymer precursor in the fluidic device, thereby forming a compartment comprising a polymer matrix wall within the fluid, wherein the fluid comprises a plurality of cells, wherein the compartment encloses a cell of the plurality of cells, and wherein the compartment physically separates the cell from an additional cell of the plurality of cells;

b) incubating the cell for at least about 1 minute, wherein the cell is inside of the compartment during the incubating; and

c) performing an assay within the compartment, wherein the assay comprises detecting a nucleic acid from the cell.

Assignments (1)

Continuity (9)

References Cited (400)