SINGLE CELL CAPTURE WITH CAPTURE CHIPS

• US 20170320038A1
• Filed: 07/27/2017
• Published: 11/09/2017
• Est. Priority Date: 06/12/2014
• Status: Active Application

• Current Assignee: Takara Bio Usa, Inc. (Ningbo Junsheng Investment Group Co., Ltd.)
• Original Assignee: Takara Bio Usa, Inc. (Ningbo Junsheng Investment Group Co., Ltd.)
• Inventors: Husain, Syed A., Griswold, Bradley L., Slater, Michael, Espinoza Vallejos, Patricio A., Dunne, Jude, Hein, Glenn, Srinivasan, Maithreyan
• Application Number:
  US15/661,528
• Publication Number:
  US 20170320038A1
• Time in Patent Office: Days
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00709 Type of synthesis
• CPC Class Codes: B01J 2219/00718 Type of compounds synthesised
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0893 having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 2300/165 Specific details about hydrophobic, oleophobic surfaces
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/50857 using arrays or bundles of open capillaries for holding samples
• CPC Class Codes: G01N 1/405 by adsorption or absorption

Abstract

The present invention provides methods, systems, assemblies, and articles for capturing single cells with a capture chip. In certain embodiments, the capture chip comprises a substrate comprising a plurality of cell-sized dimples or wells that each allow a single cell to be captured from a cell suspension. In some embodiments, the dimples or wells of the capture chip align with the holes or wells of a multi-well through-hole chip, and/or a multi-well chip, such that the cell, or the contents of the single cell, may be transferred to a corresponding well of the multi-well chip. In particular embodiments, the bottom of each dimple or well of the capture chip has a positive electrical charge sufficient to attract cells from a cell suspension flowing over the dimples or wells.

Claims

1. A system comprising:

a) a capture chip comprising a substrate comprising a plurality of cell-sized dimples or wells that each allow a single cell to be captured from a cell mixture; and

b) a multi-well through-hole chip, wherein said multi-well through-hole chip comprises a plurality of holes, and when combined with a backing or said capture chip, forms a multi-well chip which comprises a plurality of wells; and

wherein said plurality of cell-sized dimples or wells matches one-for-one, and aligns with, said plurality of holes in said multi-well through-hole chip.

2. The system of claim 1, wherein some or all of said cell-sized dimples or wells each contain a single cell, and/or wherein said plurality of wells in said multi-well chip:

i) have a volume of 50 and 5000 nl, and/or ii) comprise at least 300 wells.

3. The system of claim 1, wherein said capture chip and said multi-well through-hole chip are attached to each other thereby forming said multi-well chip, and wherein some or all of said plurality of wells in said multi-well chip contain reagents that detach and/or lyse cells.

4. The system of claim 3, wherein some or all of said cell-sized dimples or wells each contain a single cell.

5. The system of claim 1, wherein said substrate comprises photoresist.

6. The system of claim 1, wherein said substrate comprises an electrically conductive material.

7. The system of claim 6, wherein said electrically conductive material is optically transparent.

8. The system of claim 6, wherein said electrically conductive material comprises indium titanium oxide.

9. The system of claim 1, wherein said substrate comprises a non-conductive material and an electrically conductive material.

10. The system of claim 1, wherein each of said dimples or wells of said capture chip has a bottom and at least one side wall.

11. The system of claim 10, wherein said bottom is composed of a material comprising an electrically conductive material, and said at least one side wall is composed of a non-conductive material.

12. The system of claim 11, wherein said electrically conductive material is optically transparent, and wherein said capture chip comprises at least 50 dimples or wells.

13. The system of claim 11, wherein said electrically conductive material comprises indium titanium oxide.

14. The system of claim 11, wherein said at least one side wall is composed of a material comprising photoresist.

15. The system of claim 6, further comprising:

c) an electrical field generator configured to be electrically linked to said electrically conductive material.

16. A method of loading a capture-chip with cells comprising:

a) flowing a cell suspension over a capture-chip, wherein said capture-chip comprises a substrate with a plurality of cell-sized dimples or wells formed therein, and wherein said flowing causes a single cell to be captured in each of said plurality of cell-sized dimples or wells; and

b) washing said capture-chip such that excess cells from said cell suspension not captured in said plurality of dimples or wells are removed.

17. The method of claim 16, wherein said substrate comprises non-conductive material and a first electrically conductive material.

18. The method of claim 17, wherein each of said plurality of cell-sized dimples or wells comprises a bottom and at least one side.

19. The method of claim 18, wherein said bottom comprises said first electrically conductive material, and said at least one side comprises said non-conductive material.

20. The method of claim 19, wherein said first electrically conductive material is electrically linked to an electrical field generator, and wherein said electrical field generator imparts a positive charge to each of said dimple bottoms sufficient to attract cells from said cell suspension.

21. The method of claim 20, wherein said substrate further comprises a second electrically conductive material, wherein said second electrically conductive material has zero or negative voltage with respect to said first electrically conductive material.

22. The method of claim 17, wherein said electrically conductive material is optically transparent.

23. The method of claim 17, wherein said electrically conductive material comprises indium titanium oxide.

24. The method of claim 19, wherein said non-conductive material comprises photoresist, and wherein said capture-chip comprises at least 50 of said dimples.