IP Library › Granted Patent US 12,151,242
Granted Patent B2
US 12,151,242 · App. 18/593,787 · Granted Nov 26, 2024

Methods for analyzing biological samples in a fluidic device comprising generating a polymer matrix from precursors at specific locations comprising biological components

Inventors: Tarun Kumar Khurana (Palo Alto, CA); Ali Agah (Palo Alto, CA); Yir-Shyuan Wu (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
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,151,242
App. No.
18/593,787
Granted
Nov 26, 2024
Kind
B2
Abstract

Described herein are systems and methods for analyzing biological samples. Including a method for processing an analyte, comprising providing a fluidic device comprising the analyte and one or more polymer precursors; selecting a discrete area within said fluidic device; providing an energy source in optical communication with fluidic device; and selectively supplying a unit of energy generated from the energy source to the fluidic device to generate a polymer matrix within the fluidic device, wherein the polymer matrix is within the discrete area or adjacent to the discrete area.

Claims (34)

1. A method for processing a biological sample, comprising:

(a) introducing the biological sample into a fluidic device, wherein the biological sample comprises a biological component;

(b) introducing polymer precursors into the fluidic device;

(c) determining a location of the biological component within the fluidic device; and

(d) selectively polymerizing the polymer precursors to generate a polymer matrix from the polymer precursors within the fluidic device, wherein the polymer matrix at least partially encapsulates the biological component.

2. The method of claim 1 , wherein the polymer matrix forms a chamber, and wherein an interior of a portion of the chamber comprises the biological component.

3. The method of claim 1 , wherein the polymer matrix forms a hydrogel.

4. The method of claim 1 , wherein the fluidic device comprises a surface having a capture probe immobilized thereto, and wherein the capture probe couples to the biological component to immobilize the biological component to the surface.

5. The method of claim 4 , wherein the capture probe comprises one or more functional groups capable of interacting with the biological component.

6. The method of claim 5 , wherein the one or more functional groups comprise a complementary DNA sequence to target DNA or RNA of the biological component.

7. The method of claim 1 , wherein the determining of the location of the biological component within the fluidic device in (c) comprises using a detector to image the fluidic device.

8. The method of claim 7 , wherein the detector is coupled to an energy source configured to emit energy.

9. The method of claim 8 , wherein the energy source is a light generating device, and wherein the energy comprises light.

10. The method of claim 8 , wherein the energy source is in optical communication with the fluidic device.

11. The method of claim 9 , further comprising generating a virtual mask based on the location of the biological component within the fluidic device determined in (c).

12. The method of claim 11 , further comprising projecting the virtual mask using the light emitted from the light generating device.

13. The method of claim 12 , wherein the light projected from the virtual mask is used to selectively polymerize the polymer precursors to generate the polymer matrix in (d).

14. The method of claim 11 , wherein the virtual mask is generated from a spatial light modulator (SLM).

15. The method of claim 14 , wherein the SLM is a digital micromirror device (DMD).

16. The method of claim 1 , wherein the biological component comprises a cell.

17. The method of claim 16 , further comprising releasing an analyte from the cell.

18. The method of claim 17 , wherein the analyte comprises messenger ribonucleic acids (RNAs), and wherein the method further comprises capturing the messenger RNAs with one or more capture oligonucleotides located on a portion of the fluidic device encompassed by the polymer matrix.

19. The method of claim 18 , further comprising degrading the polymer matrix and loading the fluidic device with reverse transcriptase reagents to copy the captured messenger RNAs to produce complementary deoxyribonucleic acids (DNAs).

20. The method of claim 19 , further comprising sequencing the complementary DNAs.

21. The method of claim 18 , wherein the one or more capture oligonucleotides each comprise a barcode which indicates a position within the fluidic device.

22. The method of claim 1 , wherein the polymer matrix comprises polymer matrix walls having an annular-like cross-section.

23. The method of claim 1 , further comprising performing one or more functional assays on the biological component, wherein the one or more functional assays assess cell viability, cell morphology, cell secretions, cell responses, intercellular interactions, or any combination thereof.

24. The method of claim 23 , wherein the one or more functional assays are performed using bright-field phase contrast or fluorescent imaging of the biological component.

25. The method of claim 1 , further comprising, prior to (d), selecting the biological component from the biological sample based on one or more optical signals of the biological component.

26. The method of claim 1 , wherein the fluidic device comprises a channel with an inlet and an outlet, wherein the channel comprises a first surface and a second surface disposed opposite one another across the channel, wherein the polymer matrix forms a wall extending between the first surface and the second surface, thereby forming a chamber, and wherein the chamber encapsulates the biological component.

27. The method of claim 1 , wherein the polymer matrix comprises a hydrogel, and wherein the hydrogel comprises acrylamide and poly (ethylene glycol).

28. The method of claim 1 , wherein the polymer matrix comprises a degradable polymer matrix, and wherein the method further comprises degrading the polymer matrix with a degradation agent selected from the group consisting of dithiothreitol (DTT), tris (2-chloroethyl) phosphate (TCEP), tetrahydropyran (THP), and sodium periodate (NaIO4).

29. The method of claim 1 , wherein the polymer matrix comprises a photo-degradable polymer matrix, and wherein the method further comprises photo-degrading the polymer matrix by selectively projecting a light beam.

30. The method of claim 1 , wherein the introducing in (a) and the introducing in (b) are performed simultaneously.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 19, 2024
From: KHURANA, TARUN KUMAR; AGAH, ALI; WU, YIR-SHYUAN; YASAR, FILIZ GORPE
To: CELLANOME, INC.
Reel/Frame 066829/0484 →
Continuity (5)
Continuation 18219545 · Jul 7, 2023
Continuation PCTUS2022011720 · Jan 7, 2022
Provisional Application 63253500 · Oct 7, 2021
Provisional Application 63135463 · Jan 8, 2021
Related Publication 20240198332A1 · Jun 20, 2024
Cited By (7)
US 12,440,837 US 12,569,847 US 12,576,399 US 12,576,400 US 12,600,817 US 12,661,647 US 12,714,996