Flow cell device for single cell analysis, and single cell analysis device

Patent metadata

Field	Value
Publication number	US-2018304262-A1
Application number	US-201515544888-A
Country	US
Kind code	A1
Filing date	Feb 3, 2015
Priority date	Feb 3, 2015
Publication date	Oct 25, 2018
Grant date	—

Abstract

(1) Plural reaction chambers each including at least one cell capturing unit and at least one nucleic acid capturing unit connected via a flow channel, (2) a first liquid flow channel commonly connected to the plural reaction chambers on a first surface side, (3) plural second liquid flow channels connected to the plural reaction chambers in a one-on-one relationship on the first surface side, and (4) a third liquid flow channel commonly connected to the plural reaction chambers on a second surface side opposite to the first surface, are provided in a flow cell device for single cell analysis.

First claim

1 . A flow cell device for single cell analysis, comprising:
a plurality of reaction chambers each including at least one cell capturing unit and at least one nucleic acid capturing unit connected via a flow channel; a first liquid flow channel that is commonly connected to the plurality of reaction chambers on a first surface side; a plurality of second liquid flow channels that are connected to the plural reaction chambers in a one-on-one relationship on the first surface side; and a third liquid flow channel that is commonly connected to the plural reaction chambers on a second surface side opposite to the first surface.

2 . The flow cell device for single cell analysis according to claim 1 , wherein the cell capturing unit corresponds to the nucleic acid capturing unit in a one-on-one relationship.

3 . The flow cell device for single cell analysis according to claim 1 , wherein the cell capturing unit is disposed on the first surface side and the nucleic acid capturing unit is disposed on the second surface side.

4 . The flow cell device for single cell analysis according to claim 1 , wherein the nucleic acid capturing unit has a porous structure or a structure in which a large number of beads are packed.

5 . The flow cell device for single cell analysis according to claim 1 , wherein a DNA probe in the nucleic acid capturing unit includes a sequence for identifying the position at which the nucleic acid capturing unit is disposed and a capturing sequence for capturing a nucleic acid.

6 . The flow cell device for single cell analysis according to claim 1 , wherein all the inner walls of the plurality of reaction chambers have been subjected to a hydrophilic treatment and an inner wall surface of the area of the first liquid flow channel that connects the plurality of reaction chambers has been subjected to a hydrophobic treatment.

7 . A flow cell device for single cell analysis, comprising:
a plurality of reaction chambers each including at least one cell capturing unit and at least one nucleic acid capturing unit connected via a flow channel; a plurality of first liquid flow channels that are connected to the plurality of reaction chambers in a one-on-one relationship on a first surface side; and a second liquid flow channel that is commonly connected to the plurality of reaction chambers on a second surface side opposite to the first surface.

8 . The flow cell device for single cell analysis according to claim 7 , wherein the cell capturing unit corresponds to the nucleic acid capturing unit in a one-on-one relationship.

9 . The flow cell device for single cell analysis according to claim 7 , wherein the cell capturing unit is disposed on the first surface side and the nucleic acid capturing unit is disposed on the second surface side.

10 . The flow cell device for single cell analysis according to claim 7 , wherein the nucleic acid capturing unit has a porous structure or a structure in which a large number of beads are packed.

11 . The flow cell device for single cell analysis according to claim 7 , wherein a DNA probe in the nucleic acid capturing unit includes a sequence for identifying the position at which the nucleic acid capturing unit is disposed and a capturing sequence for capturing a nucleic acid.

12 . The flow cell device for single cell analysis according to claim 7 , wherein the first liquid flow channel is formed as an opening of the reaction chamber for exposing the cell capturing unit to the outside.

13 . A single cell analysis apparatus, comprising:
a stage on which a flow cell device for single cell analysis is placed, the flow cell device including, a plurality of reaction chambers each including at least one cell capturing unit and at least one nucleic acid capturing unit connected via a flow channel, a plurality first liquid flow channels connected to the plurality of reaction chambers in a one-on-one relationship on a first surface side, and a second liquid flow channel commonly connected to the plurality of reaction chambers on a second surface side opposite to the first surface; a stage controlling system for controlling the position of the stage; an introduction controller for individually controlling introduction of a first liquid into the plurality of first fluid flow channels: and a discharge controller for controlling discharge of a second liquid from the second fluid flow channel.

14 . The single cell analysis apparatus according to claim 13 , further comprising, on the first surface side of the flow cell device for single cell analysis:
a second introduction controller for introducing a third liquid into a third liquid flow channel that is commonly connected to the plurality of reaction chambers; and a second discharge controller for discharging the third liquid from the third liquid flow channel.

15 . The single cell analysis apparatus according to claim 13 , further comprising:
an optical microscope for observing the flow cell device for single cell analysis, and correlating information acquired from an observation area to positional information acquired from the stage controlling system; a DNA sequencer for analyzing a sequence of a nucleic acid amplification product obtained for each of the nucleic acid capturing units of the flow cell device for single cell analysis, and correlating the analysis result to the positional information acquired by the stage controlling system; and an information integration system for integrating the information acquired from an observation area by the optical microscope and the analysis result for each of the nucleic acid capturing units acquired by the DNA sequencer.

Assignees

• Hitachi Ltd

Inventors

• Shirai Masataka
• Sakai Tomoyuki

Classifications

• C12Q1/6869

  Methods for sequencing · CPC title

• G06F19/22

  Physics · mapped topic

• C12N15/1003

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

• B01L3/502715

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• B01L3/502761 Primary

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title