MicroRNA In Situ Hybridization in Paraffin-Embedded Human Articular Cartilage and Mouse Knee Joints

Helal Endisha; Mohit Kapoor

doi:10.1007/978-1-0716-1119-7_7

MicroRNA In Situ Hybridization in Paraffin-Embedded Human Articular Cartilage and Mouse Knee Joints

Methods Mol Biol. 2021:2245:93-103. doi: 10.1007/978-1-0716-1119-7_7.

Authors

Helal Endisha^{1

2}, Mohit Kapoor^{3

4

5}

Affiliations

¹ Arthritis Program, Krembil Research Institute, University Health Network, Toronto, ON, Canada.
² Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.
³ Arthritis Program, Krembil Research Institute, University Health Network, Toronto, ON, Canada. mohit.kapoor@uhnresearch.ca.
⁴ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada. mohit.kapoor@uhnresearch.ca.
⁵ Department of Surgery, University of Toronto, Toronto, ON, Canada. mohit.kapoor@uhnresearch.ca.

PMID: 33315197
DOI: 10.1007/978-1-0716-1119-7_7

Abstract

MicroRNA (miRNA) in situ hybridization (ISH) is a highly sensitive method that allows for the detection of expression and distribution of miRNAs in fixed paraffin-embedded tissues. MiRNA ISH requires time-consuming optimization based on the tissue type analyzed, method of tissue fixation, and miRNA detection probe. Here, we provide the optimized miRNA ISH protocol for human cartilage and mouse whole knee joints that also entails the necessary steps for sample collection, processing, and preparation for high-quality ISH staining.

Keywords: Cartilage ,; Formalin-fixed; In situ hybridization,; Locked nucleic acid,; Microrna; Mouse knee joint,; Paraffin-embedded (FFPE)..

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cartilage, Articular / metabolism*
Humans
In Situ Hybridization* / methods
Knee Joint / metabolism*
Mice
MicroRNAs / genetics*
Oligonucleotides
Paraffin Embedding
Tissue Fixation

Substances

MicroRNAs
Oligonucleotides
locked nucleic acid