In situ Hybridization (ISH) and Quantum Dots (QD) of miRNAs

Copyright © 2017 The Authors; exclusive licensee Bio-protocol LLC..

miRNA are short non-coding RNA which inhibit translation of mRNA. miRNA regulate several cellular processes. Certain miRNA are known to induce oncogenesis. miRNA can be measured by real-time PCR and be imaged using a combination of in situ hybridization (ISH) and quantum dots (QD). The advantage of using quantum dots is that several miRNA can be simultaneously measured using multiplexed QD. Additionally, miRNA can be visualized in different regions of the tissue. Since miRNA are biomarkers of various disease states, miRNA can be visualized and quantitated in tissue sections for diagnostic and prognostic purposes. Here we describe ISH-QD analysis of tissue sections. Tissue sections from xenografts or clinical specimens are used. These are deparaffinized, treated with Proteinase K and hybridized with a biotin-probe to specific to the miRNA. The in situ hybridization is performed by labeling the biotin-probes and followed by labeling with streptavidin tagged quantum dots. Image acquisition of the quantum dots is performed and analyzed for the miRNA expression levels. Combining ISH and QD gives a powerful tool to detect miRNA in different cells of the tissue.

Medienart:

E-Artikel

Erscheinungsjahr:

2017

Erschienen:

2017

Enthalten in:

Zur Gesamtaufnahme - volume:7

Enthalten in:

Bio-protocol - 7(2017), 4 vom: 20. Feb., Seite e2138

Sprache:

Englisch

Beteiligte Personen:

Josson, Sajni [VerfasserIn]
Gururajan, Murali [VerfasserIn]
W K Chung, Leland [VerfasserIn]

Links:

Volltext

Themen:

Biomarkers
Cancer
Cancer associated stroma
Cellular compartments
In situ hybridization
Journal Article
MiRNA
Multiplexed quantum dots
Tissue staining

Anmerkungen:

Date Revised 07.02.2024

published: Electronic-eCollection

Citation Status PubMed-not-MEDLINE

doi:

10.21769/BioProtoc.2138

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM330012703