IP Library › Granted Patent US 11,125,752
Granted Patent B2
US 11,125,752 · App. 16/834,569 · Granted Sep 21, 2021

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 A. Espinoza-Vallejos (Fremont, CA)
Assignee: Takara Bio USA, Inc.
G01N33/57415B01L3/5085G01N33/57492G06T7/0012B01L2300/0829B01L2300/0893B01L2300/0896C12Q1/6886C12Q2600/158G01N2333/82G06T2207/10056G06T2207/30024G06T2207/30242
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Quick Facts
Patent No.
US 11,125,752
App. No.
16/834,569
Granted
Sep 21, 2021
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 (27)

1. A method for performing analysis on single cells of interest, the method comprising:

a. labeling at least one cell in a cell suspension with a first detectable label and a second detectable label, wherein the first detectable label identifies single cells while the second detectable label identifies cells of interest in the suspension;

b. dispensing the suspension into a multi-well device using a robotic liquid dispensing system;

c. generating a dispense map using a computer processor based on signals from the first detectable label and the second detectable label; and

d. providing instructions, based on the dispense map, to the robotic liquid dispensing system to only dispense a reagent into wells containing the single cells of interest.

2. The method of claim 1 , wherein the cell suspension comprises cells at a concentration such that, on average, one cell is present in the cell suspension.

3. The method of claim 1 , wherein the detectable label comprises a cellular label.

4. The method of claim 3 , wherein the cellular label comprises a stain, an antibody, nanomaterials, or any combination thereof.

5. The method of claim 1 , wherein the first detectable label is fluorescent at a first wavelength and the second detectable label is fluorescent at a second wavelength.

6. The method of claim 1 , wherein the multi-well device comprises at least 75 wells.

7. The method of claim 6 , wherein said at least 75 wells is at least 150 wells.

8. The method of claim 1 , wherein the robotic liquid dispensing system comprises a plurality of fluidic dispensing channels, wherein the channels are configured to move between a source container with the cell suspension or the reagent and the multi-well device.

9. The method of claim 1 , wherein the reagent is selected from the group consisting of: a cell culture component, a cell differentiation reagent, a cell surface marker, an antibody, a nanomaterial, a cell lysis reagent, a drug, a nucleic acid, a lipid, a carbohydrate and a protein and combinations thereof.

10. A method for performing analysis on single cells of interest, the method comprising:

a. labeling at least one cell in a cell suspension with a first detectable label, wherein the first detectable label identifies single cells;

b. dispensing the suspension into a multi-well device using a robotic liquid dispensing system;

c. labeling at least one cell in the multi-well device with a second detectable label, wherein the second detectable label identifies cells of interest in the suspension;

d. generating a dispense map using a computer processor based on signals from the first detectable label and the second detectable label; and

e. providing instructions, based on the dispense map, to the robotic liquid dispensing system to only dispense a reagent into wells containing the single cells of interest.

11. The method of claim 10 , wherein the cell suspension comprises cells at a concentration such that, on average, one cell is present in the cell suspension.

12. The method of claim 10 , wherein the detectable label comprises a cellular label.

13. The method of claim 12 , wherein the cellular label comprises a stain, an antibody, nanomaterials, or any combination thereof.

14. The method of claim 10 , wherein the first detectable label is fluorescent at a first wavelength and the second detectable label is fluorescent at a second wavelength.

15. The method of claim 10 , wherein the multi-well device comprises at least 75 wells.

16. The method of claim 12 , wherein said at least 75 wells is at least 150 wells.

17. The method of claim 10 , wherein the robotic liquid dispensing system comprises a plurality of fluidic dispensing channels, wherein the channels are configured to move between a source container with the cell suspension or the reagent and the multi-well device.

18. The method of claim 10 , wherein the reagent is selected from the group consisting of: a cell culture component, a cell differentiation reagent, a cell surface marker, an antibody, a nanomaterial, a cell lysis reagent, a drug, a nucleic acid, a lipid, a carbohydrate and a protein and combinations thereof.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 8, 2020
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 052609/0290 →
MERGER Recorded May 8, 2020
From: WAFERGEN, INC.
To: TAKARA BIO USA, INC.
Reel/Frame 052609/0295 →
Continuity (5)
Continuation 15049056 · Feb 20, 2016
Provisional Application 62118965 · Feb 20, 2015
Provisional Application 62147756 · Apr 15, 2015
Provisional Application 62237307 · Oct 5, 2015
Related Publication 20200225236A1 · Jul 16, 2020