System for analyzing single cell

Patent metadata

Field	Value
Publication number	US-2019194645-A1
Application number	US-201716329682-A
Country	US
Kind code	A1
Filing date	Mar 6, 2017
Priority date	Sep 5, 2016
Publication date	Jun 27, 2019
Grant date	—

Abstract

A device is provided for capturing a nucleic acid in a single cell, having: a two-dimensional array comprising a substrate, a plurality of single cell capturing holes provided on one surface of the substrate, and a nucleic acid capturing region comprising, on the inside of the substrate, a nucleic acid capturing body, which is configured to capture the nucleic acids extracted from the individual cells respectively captured by the single cell capturing hole; a flow channel which is provided adjacent to the nucleic acid capturing region of the substrate, and is configured to discharge a solution in the nucleic acid capturing region; and a cylindrical structure body which is arranged on the substrate at the time of introducing a cell suspension and encloses a plurality of the single cell capturing holes, wherein the cylindrical structure body is removed after the cells are captured by the single cell capturing holes.

First claim

1 . A device for capturing a nucleic acid in a single cell, comprising:
a two-dimensional array comprising a substrate, a plurality of single cell capturing holes provided on one surface of the substrate, and a nucleic acid capturing region comprising, on the inside of the substrate, a nucleic acid capturing body, which is configured to capture the nucleic acids extracted from the individual cells respectively captured by the single cell capturing hole; a flow channel which is provided adjacent to the nucleic acid capturing region of the substrate, and is configured to discharge a solution in the nucleic acid capturing region; and a cylindrical structure body which is arranged on the substrate at the time of introducing a cell suspension and encloses a plurality of the single cell capturing holes, wherein the cylindrical structure body is removed after the cells are captured by the single cell capturing holes.

2 . The device for capturing a nucleic acid in a single cell according to claim 1 , wherein when the cylindrical structure body is arranged on the substrate, the device further comprises a means for controlling the distance between the surface on which the single cell capturing holes of the substrate are provided and the end of the cylindrical structure body.

3 . The device for capturing a nucleic acid in a single cell according to claim 2 , wherein the means is a fixing tool provided in the cylindrical structure body.

4 . The device for capturing a nucleic acid in a single cell according to claim 2 , wherein the means is a mechanism which is configured to control the movement of the cylindrical structure body.

5 . The device for capturing a nucleic acid in a single cell according to claim 1 , wherein the nucleic acid is mRNA.

6 . The device for capturing a nucleic acid in a single cell according to claim 1 , wherein the nucleic acid capturing body is magnetic beads.

7 . A system for analyzing a nucleic acid in a single cell, comprising:
a two-dimensional array comprising a substrate, a plurality of single cell capturing holes provided on one surface of the substrate, and a nucleic acid capturing region comprising, on the inside of the substrate, a nucleic acid capturing body, which is configured to capture the nucleic acids extracted from the individual cells respectively captured by the single cell capturing hole; a means for introducing a cell suspension to the surface of the substrate, on which the single cell capturing holes are provided; a flow channel which is provided adjacent to the nucleic acid capturing region of the substrate, and is configured to discharge a solution in the nucleic acid capturing region; a means for applying a negative pressure from the side opposite the side of the substrate, on which the single cell capturing holes are provided; a cylindrical structure body enclosing the plurality of single cell capturing holes; and a mechanism which is configured to arrange the cylindrical structure body on the substrate when introducing the cell suspension and to remove the cylindrical structure body after capturing the cells by the single cell capturing holes.

8 . The system for analyzing a nucleic acid in a single cell according to claim 7 , wherein when the cylindrical structure body is arranged on the substrate, the mechanism is configured to control the distance between the surface of the substrate, on which the single cell capturing holes are provided, and the end of the cylindrical structure body, or
when the cylindrical structure body is arranged on the substrate, the cylindrical structure body has a fixing tool for controlling the distance between the surface on which the single cell capturing holes of the substrate are provided and the end of the cylindrical structure body.

9 . The system for analyzing a nucleic acid in a single cell according to claim 7 , wherein the means for introducing the cell suspension is provided with a means for controlling the amount of the cell suspension to be dispensed.

10 . A method for capturing a nucleic acid in each of a single cell from a cell suspension comprising a plurality of cells, comprising:
preparing the device according to claim 1 ; introducing the cell suspension to the cylindrical structure body in a state in which the cylindrical structure body is arranged on the substrate; applying a negative pressure from the side opposite the side of the substrate, on which the single cell capturing holes are provided to capture the cells by the single cell capturing holes; removing the cylindrical structure body; and
lysing the captured cell, and capturing the nucleic acid by the nucleic acid capturing body.

11 . A method for capturing a nucleic acid in each of a single cell from a cell suspension comprising a plurality of cells, comprising:
preparing the system according to claim 7 ; introducing the cell suspension to the cylindrical structure body in a state in which the cylindrical structure body is arranged on the substrate; applying a negative pressure from the side opposite the side of the substrate, on which the single cell capturing holes are provided to capture the cells by the single cell capturing holes; removing the cylindrical structure body; and
lysing the captured cell, and capturing the nucleic acid by the nucleic acid capturing body.

Assignees

• Hitachi Ltd

Inventors

• Sakai Tomoyuki
• Shirai Masataka

Classifications

• C12M47/06

  Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material (lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003) · CPC title

• C12N15/1013 Primary

  by using magnetic beads · CPC title

• C12N15/1006

  by means of a solid support carrier, e.g. particles, polymers · CPC title

• C12N15/09

  Recombinant DNA-technology · CPC title