Method and apparatus for collecting biomolecules from specific region of tissue section

Patent metadata

Field	Value
Publication number	US-2021181076-A1
Application number	US-201816759914-A
Country	US
Kind code	A1
Filing date	Oct 3, 2018
Priority date	Nov 17, 2017
Publication date	Jun 17, 2021
Grant date	—

Abstract

Provided is, in a sample of a cell or a micro region at a position designated in a microscope image of a tissue section, an apparatus and a method for analyzing a biomolecule, which are capable of collecting and analyzing a biomolecule in a single cell or in a micro region without damaging surrounding cells.

First claim

1 . An apparatus for collecting a biomolecule from a specific region of a tissue section, comprising:
a stage on which a tissue section is fixed; a microscope system configured to acquire an optical image of the tissue section; an energy application system configured to disrupt at least a part of a specific region of the tissue section; a dispensing pipette system configured to form a droplet on the tissue section; and a computer system configured to designate the specific region on the tissue section on an optical image acquired by the microscope system, wherein a biomolecule from the specific region being disrupted is dissolved or dispersed in the droplet.

2 . The apparatus according to claim 1 , wherein the energy application system is a laser irradiation system comprising a pulsed laser, or a vibration generating device comprising a piezoelectric element.

3 . The apparatus according to claim 1 , further comprising:
a needle-shaped component for stabilizing a position of a droplet formed on the tissue section; and a mechanism configured to keep the needle-shaped component in a vicinity of a surface of the tissue section.

4 . The apparatus according to claim 1 , wherein
on the stage, a transparent plate to which the tissue section is attached is configured to be fixed, and the computer system comprises a mechanism configured to recognize a marker on the transparent plate.

5 . The apparatus according to claim 1 , wherein the specific region is a single cell or a micro region.

6 . The apparatus according to claim 1 , wherein the tissue section is a formalin-fixed paraffin-embedded (FFPE) section.

7 . The apparatus according to claim 1 , wherein the tissue section comprises a frozen section or a cultured cell, and a water-repellent thin film disposed on the frozen section or the cultured cell.

8 . A method for collecting a biomolecule from a specific region of a tissue section, comprising:
microscopically observing a tissue section; designating a specific region of the tissue section; dispensing a recovery solution on the tissue section so that a droplet to be formed covers the specific region, and forming a droplet; applying energy to at least a part of the specific region in a state where the droplet is present on the tissue section; and recovering the droplet.

9 . The method according to claim 8 , wherein applying energy comprises laser light irradiation or ultrasonic irradiation.

10 . The method according to claim 8 , wherein the droplet comprises a nucleic acid probe.

11 . The method according to claim 8 , wherein the droplet comprises a nucleic acid probe immobilized on a bead.

12 . The method according to claim 8 , wherein the droplet comprises an antibody immobilized on a bead.

13 . The method according to claim 8 , wherein the tissue section is a frozen section, and
the method further comprising: forming a water-repellent thin film on the frozen section; and forming a droplet on the water-repellent thin film.

14 . A method for collecting a biomolecule in a specific cell of a cultured cell adhered onto a substrate, comprising:
forming a water-repellent thin film on a cultured cell adhered onto a substrate; microscopically observing the cultured cell; designating a specific cell of the cultured cell; dispensing a recovery solution on the water-repellent thin film so that a droplet to be formed covers the specific cell, and forming a droplet; applying energy to at least a part of the specific cell in a state where the droplet is present on the water-repellent thin film; and recovering the droplet.

15 . The method according to claim 8 , further comprising:
collecting a biomolecule from the recovered droplet; and/or collecting a biomolecule from the recovered droplet, and amplifying and sequencing the biomolecule.

Assignees

• Hitachi Ltd

Inventors

• Shirai Masataka

Classifications

• C12N15/1003 Primary

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

• C12M33/00

  Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus (chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00) · CPC title

• C12M1/3407

  Measure of electrical or magnetical factor · CPC title

• G01N1/312 Primary

  for samples mounted on planar substrates · CPC title

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title