IP Library › Granted Patent US 12,265,088
Granted Patent B2
US 12,265,088 · App. 18/646,572 · Granted Apr 1, 2025

Systems and methods for generating droplets and performing digital analyses

Inventors: Hei Mun Christina Fan (Palo Alto, CA); Janice Hoiyi Lai (Mountain View, CA); Sixing Li (Mountain View, CA); Stephen P. A. Fodor (Palo Alto, CA); Eleen Yee Lam Shum (San Carlos, CA)
Assignee: Countable Labs, Inc.
G01N33/689C12Q1/6844G01N21/645G01N21/6486G01N2021/6482
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Quick Facts
Patent No.
US 12,265,088
App. No.
18/646,572
Filed
Apr 25, 2024
Granted
Apr 1, 2025
Kind
B2
Examiner
XU, XIAOYUN
Art Unit
1797
USPC
702/19
Abstract

This disclosure provides for devices, methods, and systems for generating a plurality of droplets within a collecting container at an extremely high rate (e.g., of at least 1 million droplets per minute, etc.), each of the plurality of droplets comprising an aqueous mixture for a digital analysis, wherein upon generation, the plurality of droplets is stabilized in position within a region of the collecting container. The inventions enable partitioning of samples for digital analyses at unprecedented rates, where readout of signals from targets within such partitions can still be achieved in accordance with various assays.

Claims (22)

1. A method, comprising:

(a) distributing a sample comprising a set of nucleic acids, across a set of partitions comprising at least 20 million partitions;

(b) amplifying the set of nucleic acids within the set of partitions;

(c) in a container, detecting signals corresponding to target nucleic acids of the set of nucleic acids, from multiple partitions of the set of partitions simultaneously, upon scanning a cross-section of the container; and

(d) counting partitions comprising signals detected in (c), wherein the counting does not comprise use of Poisson statistics associated correction factors wherein the set of partitions comprises a set of droplets, wherein the set of droplets are of an emulsion, wherein a given droplet of the set of droplets of the emulsion: (i) is in a continuous phase of the emulsion; and (ii) is surrounded by an immiscible film, wherein the immiscible film is immiscible with the continuous phase and with the given droplet of the set of droplets.

2. The method of claim 1 , wherein less than 10% of partitions of the set of partitions are occupied by nucleic acids of the set of nucleic acids.

3. The method of claim 1 , wherein the set of nucleic acids comprises cell-free nucleic acids.

4. The method of claim 1 , wherein the container has a volumetric capacity from 10 microliters to 300 microliters.

5. The method of claim 1 , wherein the emulsion comprises droplets stabilized in position in the container.

6. The method of claim 1 , wherein the partitions are stabilized in position in the container.

7. The method of claim 1 , wherein (c) comprises using a detection system for said detecting, wherein said detecting comprises optically interrogating the set of partitions using a first channel and a second channel of the detection system.

8. The method of claim 7 , wherein optically interrogating the first channel is performed within 50 seconds.

9. The method of claim 7 , wherein optically interrogating the second channel is performed within 50 seconds.

10. The method of claim 7 , wherein optically interrogating comprises performing a 3D imaging technique.

11. The method of claim 10 , wherein performing the 3D imaging technique comprises using a light-sheet imaging system.

12. The method of claim 10 , wherein performing the 3D imaging technique comprises confocal imaging.

13. The method of claim 1 , wherein (a) comprises driving the sample through a membrane.

14. The method of claim 1 , wherein the set of partitions comprises at least 30 million partitions and wherein the container has a volumetric capacity from 50 microliters through 300 microliters.

15. The method of claim 1 , wherein (c) comprises detecting signals from only a subset of partitions of the set of partitions.

16. The method of claim 1 , wherein the container comprises wells.

17. The method of claim 1 , wherein (a)-(c) are performed in less than 3 hours.

18. The method of claim 1 , wherein the sample comprises nucleic acids from single cells.

Assignments (3)
CHANGE OF NAME Recorded Apr 4, 2025
From: ENUMERIX, INC.
To: COUNTABLE LABS, INC.
Reel/Frame 070746/0051 →
CHANGE OF NAME Recorded Feb 20, 2025
From: ENUMERIX, INC.
To: COUNTABLE LABS, INC.
Reel/Frame 070287/0658 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 6, 2025
From: FAN, HEI MUN CHRISTINA; LAI, JANICE HOIYI; LI, SIXING; FODOR, STEPHEN P.A.; SHUM, ELEEN YEE LAM
To: ENUMERIX, INC.
Reel/Frame 069757/0827 →
Continuity (4)
Continuation 17711417 · Apr 1, 2022
Continuation PCTUS2022018994 · Mar 4, 2022
Provisional Application 63157292 · Mar 5, 2021
Related Publication 20240369572A1 · Nov 7, 2024
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