Granted Patent B2

US 12,576,399 · App. 19/294,044 · 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)

Assignee: Cellanome, Inc.

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

Abstract

Claims (46)

1 . A method for analyzing a cell, comprising:

a) providing energy to a fluidic device comprising the cell to polymerize a polymer precursor in the fluidic device, thereby forming a compartment comprising a polymer matrix wall, wherein the compartment at least partially encloses the cell;

b) imaging the cell, wherein the cell is located within the fluidic device, thereby generating imaging data; and

c) analyzing a nucleic acid from the imaged cell, thereby generating genomic or transcriptomic data, wherein:

the analyzing comprises capturing the nucleic acid using a capture element,

the capture element is coupled to a surface of the fluidic device,

the capture element comprises a barcode, and

the barcode is associated with a location on the surface of the fluidic device.

2 . The method of claim 1 , further comprising d) associating the genomic or transcriptomic data with the imaged cell.

3 . The method of claim 2 , wherein the associating in d) occurs after the analyzing in c), and wherein the analyzing in c) occurs after the imaging in b).

4 . The method of claim 1 , wherein:

the cell is a first cell,

the fluidic device comprises the first cell and a second cell, and

the compartment physically separates the first cell from the second cell.

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

6 . The method of claim 1 , further comprising lysing the cell, thereby releasing the nucleic acid from the cell.

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

8 . The method of claim 1 , wherein the analyzing in c) further comprises reverse transcribing the nucleic acid to form a cDNA molecule.

9 . The method of claim 8 , wherein the cDNA molecule comprises a unique molecular identifier, the barcode, a sequencing primer, or a combination thereof.

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

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

12 . The method of claim 1 , wherein the fluidic device is a microfluidic device or a nanofluidic device.

13 . The method of claim 1 , wherein the fluidic device comprises a channel, and wherein the surface is on the channel.

14 . The method of claim 4 , wherein the first cell secretes a protein, and wherein the method further 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.

15 . The method of claim 14 , wherein the capture antibody is coupled to a bead, and wherein the bead is inside of the compartment.

16 . The method of claim 4 , wherein a third cell is enclosed within the compartment with the first cell, and wherein the method further comprises analyzing an interaction between the first cell and the third cell.

17 . The method of claim 1 , wherein the polymer matrix wall extends from the surface to an additional surface of the fluidic device, wherein the surface is disposed opposite to the additional surface, thereby forming an interior of the compartment, wherein the interior of the compartment comprises the first cell.

18 . The method of claim 17 , wherein the compartment comprises a hollow cylindrical shape.

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

20 . The method of claim 19 , wherein the spatial light modulator comprises a digital micromirror device.

21 . The method of claim 19 , wherein the light is projected to a location of the first cell or adjacent to the location of the first cell.

22 . The method of claim 1 , wherein the polymer matrix wall comprises a hydrogel.

23 . The method of claim 1 , further comprising degrading at least a portion of the polymer matrix wall.

24 . The method of claim 1 , wherein the imaging comprises bright-field imaging, phase-contrast imaging, fluorescence imaging, or a combination thereof.

25 . The method of claim 4 , wherein the first 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 first cell.

26 . The method of claim 1 , wherein:

the cell is in contact with the surface of the fluidic device or an additional surface of the fluidic device,

the surface or the additional surface comprises a plurality of cell capture elements, and

subsequent to b), the cell is coupled to a cell capture element of the plurality of cell capture elements.

27 . The method of claim 26 , wherein the cell capture element comprises fibronectin, an RGD peptide, an antibody, or a combination thereof.

28 . The method of claim 1 , further comprising loading the cell into the fluidic device.

29 . The method of claim 1 , wherein the fluidic device comprises at least about 10 5 cells.

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

Assignments (1)

Continuity (8)

References Cited (400)