Device for tissue section immobilization and retention

Patent metadata

Field	Value
Publication number	US-2025067638-A1
Application number	US-202418943398-A
Country	US
Kind code	A1
Filing date	Nov 11, 2024
Priority date	Oct 25, 2021
Publication date	Feb 27, 2025
Grant date	—

Abstract

Disclosed herein, inter alia, are devices and methods for transfer and analysis of tissue sections using carrier substrates.

First claim

What is claimed is:

1 . A device comprising:
a solid support comprising a plurality of discrete tissue samples attached to the solid support, wherein each discrete tissue sample comprises: a tissue section comprising a width of about 4 mm to about 10 mm and a thickness of about 5 μm to about 12 μm; and a hydrogel substrate attached to the tissue section.

2 . The device of claim 1 , wherein each tissue section is a formalin-fixed and paraffin embedded (FFPE) tissue section.

3 . The device of claim 1 , wherein each tissue section is a frozen tissue section.

4 . The device of claim 1 , wherein a length, width, and/or diameter of each tissue section is about 4 mm to about 10 mm.

5 . The device of claim 1 , wherein a thickness of each tissue section is about 1 μm to about 20 μm.

6 . The device of claim 1 , wherein a thickness of each tissue section is less than 6 μm, 7 μm, 8 μm, 9 μm or 10 μm.

7 . The device of claim 1 , wherein each tissue section comprises breast tissue, lung tissue, colon tissue, lymph tissue, kidney tissue, bone tissue, tonsil tissue, or brain tissue.

8 . The device of claim 1 , wherein each tissue section bonds to the hydrogel substrate via a first bond having a first adhesion strength, and wherein the tissue section bonds to the solid support via a second bond having a second adhesion strength, wherein the second adhesion strength is greater than the first adhesion strength.

9 . The device of claim 1 , wherein the hydrogel substrate comprises agarose, amylose, or amylopectin.

10 . The device of claim 1 , wherein the hydrogel substrate comprises 2% to 5% agarose.

11 . The device of claim 1 wherein the hydrogel substrate comprises 3% to 20% acrylamide.

12 . The device of claim 1 , wherein the solid support is glass.

13 . A method of detecting nucleic acids in a tissue section, comprising:
contacting the device of claim 1 with a plurality of circularizable probes, wherein a circularizable probe binds to a nucleic acid molecule in the tissue section; forming a circular oligonucleotide in the tissue section, and amplifying the circular oligonucleotide to form amplification products; and detecting the amplification products by binding a fluorescently labeled probe to the amplification product and detecting the fluorescent label.

14 . The method of claim 13 , wherein prior to contacting the device with a plurality of circularizable probes, the hydrogel substrate is removed.

15 . The method of claim 13 , wherein the nucleic acid molecule is attached to a specific binding agent.

16 . The method of claim 13 , wherein the nucleic acid molecule is an RNA molecule.

17 . The method of claim 13 , wherein the nucleic acid molecule is a cDNA molecule.

18 . The method of claim 13 , where the tissue section comprises is liver tissue, kidney tissue, lung tissue, thymus tissue, adrenal tissue, skin tissue, bladder tissue, colon tissue, spleen tissue, or brain tissue.

19 . The method of claim 13 , wherein the fluorescently labeled probe is a fluorescently labeled nucleotide.

20 . The method of claim 13 , wherein the fluorescently labeled probe is a fluorescently labeled oligonucleotide.

Assignees

• Singular Genomics Systems Inc

Inventors

• Ishitsuka Yuji
• Hong Zhenmin
• Glezer Eli N
• Cang Hu
• Dempsey William
• Ding Weiqiao
• Koh Jaekyung
• Varnosfaderani Mohammad Vatankhah

Classifications

• C12N1/08

  Reducing the nucleic acid content · CPC title

• G01N33/582

  with fluorescent label · CPC title

• C12Q1/6844

  Nucleic acid amplification reactions · CPC title

• C12Q1/6834

  Enzymatic or biochemical coupling of nucleic acids to a solid phase · CPC title

• C12Q1/6825

  Nucleic acid detection involving sensors · CPC title