IP Library › Granted Patent US 12,661,647
Granted Patent B2
US 12,661,647 · App. 19/278,614 · Granted Jun 23, 2026

Methods for enclosing and analyzing a cell in a fluidic device

Inventors: Tarun Kumar Khurana (Palo Alto, CA); Ali Agah (Palo Alto, CA); Yir-Shyuan Wu (Palo Alto, CA); Pier Federico Gherardini (Palo Alto, CA); Filiz Gorpe Yasar (Palo Alto, CA)
Assignee: Cellanome, Inc.
B01L3/502715B01L3/502707B01L3/502761C12Q1/6869C12Q1/6874G01N33/54386G01N33/6842B01L2200/0647B01L2200/12B01L2300/0663B01L2300/0883B01L2300/12B01L2300/163B01L2300/18C12Q2600/158G01N2570/00
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Quick Facts
Patent No.
US 12,661,647
App. No.
19/278,614
Filed
Jul 23, 2025
Granted
Jun 23, 2026
Kind
B2
Art Unit
1681
USPC
435/6.11
Abstract

Described herein are systems and methods for analyzing biological samples (e.g., cells), including a method comprising (a) introducing a first cell into a fluidic device, (b) introducing a second cell into the fluidic device, (c) introducing a polymer precursor into the fluidic device, (d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device.

Claims (99)

1 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device; and

(e) measuring cytotoxicity of the first cell against the second cell.

2 . The method of claim 1 , wherein in (b), a plurality of second cells are introduced into the fluidic device.

3 . The method of claim 2 , wherein the measuring the cytotoxicity comprises counting dead cells, viable cells, or a combination thereof, from among the plurality of second cells.

4 . The method of claim 3 , wherein the counting comprises staining the dead cells with a vital dye.

5 . The method of claim 1 , wherein in (d), the first cell and the second cell are selectively enclosed within a chamber comprising a polymer matrix wall.

6 . The method of claim 5 , wherein the polymer matrix wall extends from a first surface of the fluidic device to a second surface opposite of the first surface, thereby forming an interior of the chamber, wherein the interior of the chamber comprises the first cell and the second cell.

7 . The method of claim 1 , further comprising detecting a protein secreted by the first cell, the second cell, or the first cell and the second cell.

8 . The method of claim 7 , wherein the detecting comprises binding the protein to a protein affinity reagent coupled to a protein-capture surface.

9 . The method of claim 8 , wherein the protein-capture surface comprises a bead.

10 . The method of claim 8 , wherein in (d), the first cell and the second cell are selectively enclosed within a chamber comprising a polymer matrix wall, and wherein the protein-capture surface is disposed within the chamber.

11 . The method of claim 7 , wherein the detecting further comprises binding a protein detection antibody to the protein, and detecting the protein detection antibody.

12 . The method of claim 1 , wherein the introducing of the first cell in (a), the introducing of the second cell in (b), and the introducing of the polymer precursor in (c) are performed at the same time.

13 . The method of claim 7 , wherein the protein is secreted by the first cell, and wherein the protein is a cytokine or an immune active protein.

14 . The method of claim 13 , wherein the protein secreted by the first cell is an interferon-gamma (IFN-g), an interferon-alpha (IFN-a), an interleukin, a colony stimulating factor (CSF), a tumor necrosis factor (TNF), or an effector molecule.

15 . The method of claim 5 , further comprising

introducing a plurality of protein-capture surfaces each coupled to a plurality of protein affinity reagents into the fluidic device;

co-enclosing at least a protein-capture surface of the plurality of protein-capture surfaces in the chamber, wherein the plurality of protein affinity reagents are configured to bind to a plurality of proteins secreted by the first cell; and

measuring binding of the plurality of proteins secreted by the first cell to the plurality of protein affinity reagents, thereby detecting the plurality of proteins secreted by the first cell.

16 . The method of claim 15 , wherein each protein-capture surface of the plurality of protein-capture surfaces comprises a single type of protein affinity reagent of the plurality of protein affinity reagents.

17 . The method of claim 1 , further comprising detecting a surface protein expressed by the first cell, the second cell, or the first cell and the second cell.

18 . The method of claim 17 , wherein the detecting comprises binding an antibody to the surface protein, and detecting the antibody.

19 . The method of claim 1 , further comprising measuring a proliferative capacity, proliferation rate, activation status, cellular identity, purity, gene expression profile, transcriptome, epigenetic profile, sequence copy number, integrated viral copy number, plasmid copy number, gene copy number, or any combination thereof, of the first cell.

20 . The method of claim 1 , wherein the measuring of the cytotoxicity comprises incubating the first cell with a dye configured to generate an optical signal in response to a characteristic of a dead cell.

21 . The method of claim 1 , further comprising detecting activation of the first cell.

22 . The method of claim 1 , further comprising identifying the first cell.

23 . The method of claim 22 , wherein the identifying comprises detecting a surface protein expressed by the first cell, detecting a protein secreted by the first cell, detecting an mRNA transcript expressed by the first cell, or any combination thereof.

24 . The method of claim 5 , further comprising removing an additional cell that is not enclosed by the chamber from the fluidic device.

25 . The method of claim 5 , wherein the polymer matrix wall comprises a hydrogel.

26 . The method of claim 5 , further comprising at least partially degrading the chamber.

27 . The method of claim 1 , further comprising, prior to (d), determining a location of at least the first cell within the fluidic device.

28 . The method of claim 27 , wherein the determining of the location is performed using a detector.

29 . The method of claim 1 , wherein the virtual photomask is generated from a spatial light modulator (SLM).

30 . The method of claim 1 , wherein the first cell comprises an effector cell, and wherein the second cell comprises a target cell.

31 . The method of claim 30 , wherein the effector cell is an immune cell.

32 . The method of claim 31 , wherein the immune cell is a cytotoxic T lymphocyte, a regulatory T cell, a CD4+ T cell, a CD8+ T cell, a natural killer cell, an antigen-presenting cell, or a dendritic cell.

33 . The method of claim 30 , wherein the target cell comprises a cancer cell.

34 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device; and

(e) detecting a protein secreted by the first cell, the second cell, or the first cell and the second cell, wherein the detecting comprises binding the protein to a protein affinity reagent coupled to a protein-capture surface, and wherein in (d), the first cell and the second cell are selectively enclosed within a chamber comprising a polymer matrix wall, and wherein the protein-capture surface is disposed within the chamber.

35 . The method of claim 34 , wherein the protein-capture surface comprises a bead.

36 . The method of claim 34 , wherein the protein is secreted by the first cell, and wherein the protein is a cytokine or an immune active protein.

37 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device; and

(e) detecting a protein secreted by the first cell, wherein the protein secreted by the first cell is an interferon-gamma (IFN-g), an interferon-alpha (IFN-a), an interleukin, a colony stimulating factor (CSF), a tumor necrosis factor (TNF), or an effector molecule.

38 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device; and

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device,

wherein the introducing of the first cell in (a), the introducing of the second cell in (b), and the introducing of the polymer precursor in (c) are performed at the same time.

39 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) introducing a plurality of protein-capture surfaces each coupled to a plurality of protein affinity reagents into the fluidic device, wherein the plurality of protein affinity reagents are configured to bind to a plurality of proteins secreted by the first cell;

(e) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively co-enclosing (i) the first cell, (ii) the second cell, and (iii) at least a protein-capture surface of the plurality of protein-capture surfaces in a chamber comprising a polymer matrix wall in the fluidic device; and

(f) measuring binding of the plurality of proteins secreted by the first cell to the plurality of protein affinity reagents, thereby detecting the plurality of proteins secreted by the first cell.

40 . The method of claim 39 , wherein each protein-capture surface of the plurality of protein-capture surfaces comprises a single type of protein affinity reagent of the plurality of protein affinity reagents.

41 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device; and

(e) detecting activation of the first cell.

42 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in a chamber comprising a polymer matrix wall in the fluidic device; and

(e) removing an additional cell that is not enclosed by the chamber from the fluidic device.

43 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device; and

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in a chamber comprising a polymer matrix wall in the fluidic device, wherein the polymer matrix wall comprises a hydrogel.

44 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device; and

(d) using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in a chamber comprising a polymer matrix wall in the fluidic device; and

(e) at least partially degrading the chamber.

45 . A method, comprising:

(a) introducing a first cell into a fluidic device;

(b) introducing a second cell into the fluidic device;

(c) introducing a polymer precursor into the fluidic device;

(d) determining a location of at least the first cell within the fluidic device; and

(e) subsequent to (d), using a virtual photomask to selectively apply light to the fluidic device to polymerize the polymer precursor, thereby selectively enclosing the first cell and the second cell in the fluidic device.

46 . The method of claim 45 , wherein the determining of the location is performed using a detector.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 23, 2026
From: KHURANA, TARUN KUMAR; AGAH, ALI; WU, YIR-SHYUAN; YASAR, FILIZ GORPE; GHERARDINI, PIER FEDERICO
To: CELLANOME. INC.
Reel/Frame 074449/0932 →
Continuity (4)
Continuation 18891555 · Sep 20, 2024
Continuation PCTUS2023015806 · Mar 21, 2023
Provisional Application 63322601 · Mar 22, 2022
Related Publication 20250345792A1 · Nov 13, 2025
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