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

Patent metadata

Field	Value
Publication number	US-11454577-B2
Application number	US-201816759914-A
Country	US
Kind code	B2
Filing date	Oct 3, 2018
Priority date	Nov 17, 2017
Publication date	Sep 27, 2022
Grant date	Sep 27, 2022

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

The invention claimed is:

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 the 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

• C12M1/3407

  Measure of electrical or magnetical factor · CPC title

• G01N2001/045

  Laser ablation; Microwave vaporisation · CPC title

• G01N1/04

  in the solid state, e.g. by cutting · CPC title

• G01N33/5308

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