Cell analysis device, apparatus, and cell analysis method using same

Patent metadata

Field	Value
Publication number	US-10752941-B2
Application number	US-201515775408-A
Country	US
Kind code	B2
Filing date	Dec 1, 2015
Priority date	Dec 1, 2015
Publication date	Aug 25, 2020
Grant date	Aug 25, 2020

Abstract

The purpose of the present invention is to provide a single cell analysis device in which the improvement of the nucleic acid capturing efficiency and the improvement of the cell capturing efficiency are both achieved and a highly accurate single cell analysis data is thereby obtained. The present invention relates to an improvement of a cell analysis device including a two-dimensional array chip having a plurality of cell capture parts capable of capturing a single cell in each of the capture parts, and nucleic acid capture parts corresponding to the respective cell capture parts, the nucleic acid capture parts being capable of capturing a nucleic acid extracted from the cell captured by the cell capture part.

First claim

The invention claimed is:

1. A cell analysis method using a cell analysis device which includes:
a solution introduction channel,
a two-dimensional array chip disposed above the solution introduction channel and having a plurality of cell capture parts and a plurality of nucleic acid capture parts, the cell capture parts being provided in communication with the solution introduction channel and each being capable of capturing a single cell, and each of the nucleic acid capture parts being provided in communication with a corresponding cell capture part of the plurality of cell capture parts and having a porous material or beads on which a nucleic acid probe configured to capture nucleic acid extracted from the single cell having been captured by the corresponding cell capture part, and
a discharge channel disposed above the two-dimensional array chip in communication with the plurality of cell capture parts and having a three-dimensional porous member,
the cell analysis method comprising:
introducing a solution containing cells to the solution introduction channel;
applying a negative pressure to the discharge channel to suck the solution from the solution introduction channel and adsorb the cells on the cell capture parts, the negative pressure bringing the two-dimensional array chip into contact with the three-dimensional porous member of the discharge channel;
maintaining the negative pressure while extracting and capturing nucleic acid with the nucleic acid probe in the nucleic acid capture parts;
introducing a reagent for nucleic acid amplification to the solution introduction channel;
applying a negative pressure to the discharge channel to suck the reagent from the solution introduction channel and introduce the reagent to the nucleic acid capture parts, the negative pressure bringing the two-dimensional array chip into contact with the three-dimensional porous member of the discharge channel;
separating the two-dimensional array chip from the three-dimensional porous member; and
after the two-dimensional array chip is separated from the three-dimensional porous member, starting an amplification reaction with the reagent and the captured nucleic acid.

2. The cell analysis method according to claim 1 , wherein the two-dimensional array chip is separated from the three-dimensional porous member by stopping application of the negative pressure to the discharge channel.

3. The cell analysis method according to claim 1 , wherein the two-dimensional array chip includes an ultrafiltration membrane or a gel membrane is disposed above the nucleic acid capture parts and has a pore size smaller than a molecular size of a product of the amplification reaction.

4. The cell analysis method according to claim 1 , wherein the two-dimensional array chip is separated from the three-dimensional porous member by introducing a separation solvent or air between the two-dimensional array chip and the three-dimensional porous member.

5. The cell analysis method according to claim 1 , wherein the two-dimensional array chip is separated from the three-dimensional porous member by an actuator connected to the three-dimensional porous member.

6. The cell analysis method according to claim 1 , further comprising:
collecting a product of the amplification reaction from the introduction channel by suction.

7. The cell analysis method according to claim 1 , wherein the applied negative pressure sucks the solution upwards counter to gravity from the solution introduction channel into the discharge channel.

8. The cell analysis method according to claim 1 , wherein the applied negative pressure to sucks the reagent upwards counter to gravity from the solution introduction channel into the discharge channel.

Assignees

• Hitachi High Tech Corp
• Hitachi High-Tech Corp

Inventors

• Shirai Masataka
• Sakai Tomoyuki
• Uchida Kenko

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6809

  Methods for determination or identification of nucleic acids involving differential detection · CPC title

• C12N15/1003

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• C12Q1/68

  involving nucleic acids · CPC title

• C40B40/08

  Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries · CPC title