IP Library › Granted Patent US 12,613,244
Granted Patent B2
US 12,613,244 · App. 17/403,489 · Granted Apr 28, 2026

Method for rapid accurate dispensing, visualization and analysis of single cells

Inventors: Alain-Albert Mir (Redwood City, CA); Thomas D. Schaal (San Francisco, CA); Chun-Wah Lin (Fremont, CA); Shanavaz Loharasp D. Nasarabadi (Fremont, CA); Jude Dunne (Menlo Park, CA); Maithreyan Srinivasan (Palo Alto, CA); Patricio Espinoza Vallejos (Fremont, CA)
Assignee: Takara Bio USA, Inc.
G01N33/57515B01L3/5085G01N33/5759G06T7/0012B01L2300/0829B01L2300/0893B01L2300/0896C12Q1/6886C12Q2600/158G01N2333/82G06T2207/10056G06T2207/30024G06T2207/30242
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Quick Facts
Patent No.
US 12,613,244
App. No.
17/403,489
Granted
Apr 28, 2026
Kind
B2
Abstract

The present disclosure provides methods, device, assemblies, and systems for dispensing and visualizing single cells. For example, provided herein are systems and methods for dispensing a dispense volume into a plurality of wells of a multi-well device, where, on average, a pre-determined number of cells (e.g., 1-20) are present in the dispense volume, and determining, via a cellular label, the number of cells present in each of the plurality of wells. Such dispensing and cell detection may be repeated a number of times with respect to wells identified as having less than the pre-determined number of cells in order increase the number wells in the multi-well device containing the desired number (e.g., a single cell).

Claims (28)

1 . A system comprising:

a) a multi-well device comprising a plurality of wells;

b) a liquid dispensing system configured to dispense a dispense volume into each of said plurality of wells; and

c) a dispense map file that provides instructions to said liquid dispensing system for dispensing liquid into wells of said multi-well device that contain single cells.

2 . The system according to claim 1 , wherein the system further comprises an image capturing system.

3 . The system according to claim 2 , wherein the image capturing system comprises an image capture device, a computer comprising a computer processor, computer memory, and image analysis software configured to generate the dispense map file from the captured image.

4 . The system according to claim 1 , wherein the system further comprises a cell suspension.

5 . The system according to claim 1 , further comprising a source of one or more reagents.

6 . The system according to claim 5 , wherein the one or more reagents are selected from the group consisting of: cell culture components, cell health measuring reagents, cell differentiation reagents, drugs, WTA reagents, WGA reagents, nucleic acid reagents, PCR reagents, reverse transcription reagents, proximity ligation assay (PLA) reagents, cell surface markers, labeled antibodies, unlabeled antibodies, detection reagents associated with antibody detection, nanomaterials, reagents for lysing cells, reagents for detection and/or resolution of nucleic acids or lipids or carbohydrates or protein cell components, and combinations thereof.

7 . A system comprising:

a) a multi-well device comprising a plurality of wells;

b) a liquid dispensing system configured to dispense liquid into wells of the plurality of wells;

c) an image capturing system; and

d) a dispense map file based on an image captured by the image capture system that provides instructions to the liquid dispensing system to only dispense a reagent into wells containing only one cell.

8 . The system according to claim 7 , wherein the plurality of wells comprises 100 or more wells.

9 . The system according to claim 8 , wherein the plurality of wells comprises 1000 or more wells.

10 . The system according to claim 7 , wherein the multi-well device comprises a multi-well chip.

11 . The system according to claim 7 , wherein the image capturing system further comprises a computer comprising a computer processor, computer memory, and image analysis software configured to generate the dispense map file from the captured image.

12 . The system according to claim 7 , wherein the system further comprises a cell suspension.

13 . The system according to claim 7 , further comprising a source of one or more reagents.

14 . The system according to claim 13 , wherein the one or more reagents are NGS reagents.

15 . The system according to claim 13 , wherein the one or more reagents are selected from the group consisting of: a primer, a template switch oligonucleotide, a polymerase, a reverse transcriptase, dNTPs, and combinations thereof.

16 . The system according to claim 13 , wherein the one or more reagents comprise a reagent comprising a barcode.

17 . The system according to claim 16 , wherein the system further comprises a cell suspension.

18 . A method comprising:

dispensing, with a liquid dispensing system, a reagent into wells of a multi-well device using a dispense map that provides instructions to the liquid dispensing system to dispense the reagent into wells containing only one cell.

19 . The method according to claim 18 , wherein the method is a method of next generation sequencing (NGS).

20 . The method according to claim 19 , wherein the reagent is selected from the group consisting of: a primer, a template switch oligonucleotide, a polymerase, a reverse transcriptase, dNTPs, and combinations thereof.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 16, 2021
From: MIR, ALAIN-ALBERT; SCHAAL, THOMAS D.; LIN, CHUN-WAH; NASARABADI, SHANAVAZ LOHARASP D.; DUNNE, JUDE; SRINIVASAN, MAITHREYAN; ESPINOZA-VALLEJOS, PATRICIO A.
To: WAFERGEN, INC.
Reel/Frame 057194/0049 →
MERGER Recorded Aug 16, 2021
From: WAFERGEN, INC.
To: TAKARA BIO USA, INC.
Reel/Frame 057194/0061 →
Continuity (6)
Continuation 16834569 · Mar 30, 2020
Continuation 15049056 · Feb 20, 2016
Provisional Application 62237307 · Oct 5, 2015
Provisional Application 62147756 · Apr 15, 2015
Provisional Application 62118965 · Feb 20, 2015
Related Publication 20210382056A1 · Dec 9, 2021
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