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In-situ Hybridization

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

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Related Experiment Video

Updated: Jul 18, 2026

A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract

A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract

Published on: August 19, 2011

Micro-capillary tube in situ hybridisation: a novel method for processing small individual samples.

A A Avilion¹, D M Bell, R Lovell-Badge

¹MRC, National Institute for Medical Research, London, United Kingdom. aavilio@nimr.mrc.ac.uk

Genesis (New York, N.Y. : 2000)

|July 13, 2000

Summary

Researchers developed a new RNA in situ hybridization method for analyzing many small tissue samples efficiently. This technique allows simultaneous analysis of 18 samples with low background, aiding gene expression studies.

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MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

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Last Updated: Jul 18, 2026

A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract

A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract

Published on: August 19, 2011

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A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons

Published on: November 14, 2018

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method

Published on: October 7, 2025

Area of Science:

Molecular Biology
Developmental Biology
Genetics

Background:

Analyzing small tissue samples for gene expression is challenging.
RNA in situ hybridization is a key technique for studying gene expression patterns.
High-throughput analysis of numerous small samples is needed.

Purpose of the Study:

To develop a novel strategy for the high-throughput analysis of small tissue samples using RNA in situ hybridization.
To enable simultaneous analysis of multiple samples with reduced background noise.
To validate the technique for studying specific gene expression in embryonic tissues.

Main Methods:

Development of a protocol for processing small tissue samples (approx. 0.4 mm x 0.5 mm) in capillary tubes.
Utilizing nylon mesh and glass beads within capillary tubes for sample processing.
Implementing standard RNA in situ hybridization protocols for simultaneous analysis of up to 18 samples.

Main Results:

Successful individual analysis of large numbers of small tissue samples.
Achieved simultaneous assaying of 18 samples without loss and with low background.
Demonstrated the technique's efficacy by examining mouse Sox2 RNA expression in embryonic chorion.
Confirmed the method's suitability for double RNA labeling and potential for protein antibody staining.

Conclusions:

The developed strategy provides an efficient method for analyzing numerous small tissue samples via RNA in situ hybridization.
This technique offers a robust platform for gene expression studies, including double labeling and potential protein analysis.
The method is particularly useful for analyzing limited or precious biological samples, such as early-stage embryos.