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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...

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Related Experiment Video

Updated: Jun 4, 2026

In Situ Hybridization for the Precise Localization of Transcripts in Plants

In Situ Hybridization for the Precise Localization of Transcripts in Plants

Published on: November 23, 2011

In Situ Hybridization to Localize mRNAs.

¹In Situ Hybridisation Service, Imperial Cancer Research Fund, London, UK.

Methods in Molecular Medicine

|February 23, 2011

Summary

This summary is machine-generated.

In situ hybridization (ISH) using 35S-labeled riboprobes offers a reliable and cost-effective method for localizing mRNA expression in pathology specimens. This isotopic technique provides reproducible results for gene expression studies in tissues.

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Last Updated: Jun 4, 2026

In Situ Hybridization for the Precise Localization of Transcripts in Plants

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Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

Published on: April 24, 2019

Area of Science:

Molecular Biology
Pathology
Genetics

Background:

In situ hybridization (ISH) is a powerful technique for localizing mRNA expression in biological tissues.
ISH provides critical gene expression data when specific antibodies are unavailable.
Human pathology specimens and experimental tissues are commonly studied using ISH.

Purpose of the Study:

To describe a preferred, reliable, and cost-effective isotopic in situ hybridization (ISH) method for routine pathology specimens.
To highlight the advantages of using 35S-labeled riboprobes for screening mRNA expression.
To present a protocol yielding reproducible and interpretable results across various cell types.

Main Methods:

Utilized formalin-fixed, paraffin-embedded tissues, including routine surgical specimens and archival hospital samples.
Employed a refined isotopic in situ hybridization (ISH) protocol using 35S-labeled riboprobes.
Focused on optimizing steps for specificity, sensitivity, and cost-effectiveness.

Main Results:

The ICRF In Situ Hybridization Service successfully hybridized over 44,000 tissue sections with a high success rate.
The 35S-labeled riboprobe method demonstrated superior reproducibility and interpretability compared to non-isotopic ISH.
The protocol achieved good results for most routine surgical specimens and useful data from archival samples.

Conclusions:

Isotopic ISH with 35S-labeled riboprobes is a valuable, cost-effective, and reliable method for mRNA localization in pathology.
This technique offers a balance of specificity, sensitivity, and reproducibility crucial for gene expression studies.
The described protocol is suitable for analyzing gene expression in a wide range of routinely processed tissue samples.