公开(公告)号：US11214796B2

公开(公告)日：2022-01-04

申请号：US15621316

申请日：2017-06-13

Applicant: HITACHI, LTD.

Inventor： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi

IPC: C12N15/10 ,  G01N33/543 ,  C12Q1/6809 ,  C12Q1/6837 ,  C40B40/08 ,  C40B50/06

Abstract: The present invention provides a method and/or means for collecting and analyzing an individual cell in a tissue, and at the same time, quantitatively monitoring the expression levels of various genes while keeping two-dimensional information in the tissue. Specifically, the present invention provides a method comprising preparing a cDNA library from mRNA while keeping two-dimensional cellular distribution information and obtaining the gene expression levels at any site or all sites at a level of single cell. More specifically, the present invention provides a method comprising preparing a cDNA library in a sheet-form from mRNA while keeping two-dimensional cellular distribution information and repeatedly using the cDNA library in the detection of the gene expression, thereby allowing measurement of the expression distribution for a number of genes at a high accuracy.

公开(公告)号：US10030240B2

公开(公告)日：2018-07-24

申请号：US14771339

申请日：2013-03-12

Applicant: Hitachi, Ltd.

Inventor： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi ,  Maiko Tanabe

IPC: C12N15/10 ,  C12Q1/6874 ,  B01L3/00 ,  B01L7/00

Abstract: In order to conduct gene expression analysis of a number of genes in a number of cells, it has been necessary to separate cells, extract genes therefrom, amplify nucleic acids, and perform sequence analysis. However, separation of cells imposes damages on the cells, and it requires the use of an expensive system. Gene expression analysis in each cell can be carried out with high accuracy by arranging a pair of structures comprising a cell trapping section and a nucleic acid trapping section in a vertical direction to extract individual genes in relevant cells, synthesizing cDNA in the nucleic acid trapping section, amplifying nucleic acids, and analyzing the sequences using a next-generation sequencer.

公开(公告)号：US11680286B2

公开(公告)日：2023-06-20

申请号：US17323311

申请日：2021-05-18

Applicant: Hitachi, Ltd.

Inventor： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi ,  Maiko Tanabe

IPC: C12Q1/6837 ,  C12N15/10

CPC classification number: C12Q1/6837 ,  C12N15/1096 ,  C12Q1/6837 ,  C12Q2525/155 ,  C12Q2565/537 ,  C12Q2565/601

Abstract: The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.

公开(公告)号：US20170283796A1

公开(公告)日：2017-10-05

申请号：US15621316

申请日：2017-06-13

Applicant: HITACHI, LTD.

Inventor： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi

IPC: C12N15/10 ,  G01N33/543 ,  C40B50/06 ,  C12Q1/68 ,  C40B40/08

CPC classification number: C12N15/1093 ,  C12Q1/6809 ,  C12Q1/6837 ,  C40B40/08 ,  C40B50/06 ,  G01N33/54306 ,  G01N2458/10 ,  C12Q2531/125 ,  C12Q2533/107 ,  C12Q2565/537

Abstract: The present invention provides a method and/or means for collecting and analyzing an individual cell in a tissue, and at the same time, quantitatively monitoring the expression levels of various genes while keeping two-dimensional information in the tissue. Specifically, the present invention provides a method comprising preparing a cDNA library from mRNA while keeping two-dimensional cellular distribution information and obtaining the gene expression levels at any site or all sites at a level of single cell. More specifically, the present invention provides a method comprising preparing a cDNA library in a sheet-form from mRNA while keeping two-dimensional cellular distribution information and repeatedly using the cDNA library in the detection of the gene expression, thereby allowing measurement of the expression distribution for a number of genes at a high accuracy.