Single cell capture with capture chips

• US 9,757,707 B2
• Filed: 10/28/2015
• Issued: 09/12/2017
• Est. Priority Date: 06/12/2014
• Status: Active Grant

• 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
• Primary Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US14/925,654
• Publication Number:
  US 20160045884A1
• Time in Patent Office: 685 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,

wherein said substrate comprises;

i) a base support layer, ii) an electrically conductive layer on top of said base support layer, and iii) a non-conductive layer substantially covering said electrically conductive layer,

wherein said substrate has at least 300 cell-sized dimples or wells formed therein that each are sized to allow only one lymphocyte sized cell to be captured from a cell mixture of lymphocyte sized cells, and

wherein each of said dimples or wells has a bottom formed from said electrically conductive layer and at least one side wall formed from said non-conductive layer;

b) a flow cell comprising at least one flow channel, wherein said flow cell is structured so as to be mated with said capture chip to form a water-tight assembly, wherein said water-tight assembly allows said cell mixture to move over said capture chip in said at least one flow channel; and

c) an electrical field generator that is, or configured to be, electrically linked to said electrically conductive layer, wherein said electrical field generator imparts a positive charge to each of said dimple or well bottoms sufficient to attract a cell from said cell mixture when flowing over said dimples or wells in said flow cell.

2. The system of claim 1, further comprising d) a plurality of single lymphocyte cells, wherein some or all of said cell-sized dimples or wells each contain only one of said lymphocyte cells.

3. The system of claim 1, wherein said non-conductive layer comprises photoresist.

4. The system of claim 1, wherein said electrically conductive layer is optically transparent.

5. The system of claim 1, wherein said electrically conductive layer comprises indium titanium oxide.

6. The system of claim 1, wherein said flow cell is mated with said capture chip to form said water-tight assembly.

7. The system of claim 1, wherein said at least one side wall is composed of a material comprising photoresist.

8. A system comprising:

a) a capture chip comprising a substrate,

wherein said substrate comprises;

i) a base support layer, ii) an electrically conductive layer on top of said base support layer, and iii) a non-conductive layer substantially covering said electrically conductive layer,

wherein said substrate has at least 300 cell-sized dimples or wells formed therein that each are sized to allow only one fibroblast sized cell to be captured from a cell mixture of fibroblast sized cells, and

wherein each of said dimples or wells has a bottom formed from said electrically conductive layer and at least one side wall formed from said non-conductive layer;

b) a flow cell comprising at least one flow channel, wherein said flow cell is structured so as to be mated with said capture chip to form a water-tight assembly, wherein said water-tight assembly allows said cell mixture to move over said capture chip in said at least one flow channel; and

c) an electrical field generator that is, or configured to be, electrically linked to said electrically conductive layer, wherein said electrical field generator imparts a positive charge to each of said dimple or well bottoms sufficient to attract a cell from said cell mixture when flowing over said dimples or wells in said flow cell.

9. The system of claim 8, wherein said flow cell is mated with said capture chip to form said water-tight assembly.

10. A system comprising:

a) a capture chip comprising a substrate,

wherein said substrate comprises;

i) a base support layer, ii) an electrically conductive layer on top of said base support layer, and iii) a non-conductive layer substantially covering said electrically conductive layer,

wherein said substrate has at least 300 cell-sized dimples or wells formed therein that each are sized to allow only one hematopoietic stem cell sized cell to be captured from a cell mixture of hematopoietic stem cell sized cells, and

wherein each of said dimples or wells has a bottom formed from said electrically conductive layer and at least one side wall formed from said non-conductive layer;

b) a flow cell comprising at least one flow channel, wherein said flow cell is structured so as to be mated with said capture chip to form a water-tight assembly, wherein said water-tight assembly allows said cell mixture to move over said capture chip in said at least one flow channel; and

c) an electrical field generator that is, or configured to be, electrically linked to said electrically conductive layer, wherein said electrical field generator imparts a positive charge to each of said dimple or well bottoms sufficient to attract a cell from said cell mixture when flowing over said dimples or wells in said flow cell.

11. The system of claim 10, wherein said flow cell is mated with said capture chip to form said water-tight assembly.