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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,...

Southern Blot

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...

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Related Experiment Video

Updated: May 1, 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

Developments in in situ hybridisation.

Andrew Cassidy¹, Julia Jones¹

¹Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Cambridge, United Kingdom.

Methods (San Diego, Calif.)

|April 22, 2014

Summary

This summary is machine-generated.

In situ hybridisation (ISH) visualizes specific nucleic acids in tissues. This review focuses on messenger RNA (mRNA) and micro RNA (miRNA) detection in FFPE tissues using non-radioactive methods.

Keywords:

Branched DNA amplification In situ hybridisation Locked nucleic acid Rolling circle amplification Signal amplification Tissue fixation

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In Situ Hybridization for the Precise Localization of Transcripts in Plants

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Area of Science:

Molecular Biology
Histology
Genetics

Background:

In situ hybridisation (ISH) is a powerful technique for detecting specific DNA or RNA sequences within their native cellular or tissue context.
It is widely used in research and diagnostics for visualizing gene expression, RNA localization, and genomic alterations.
The method has evolved significantly over decades, with numerous refinements enhancing its sensitivity and applicability.

Purpose of the Study:

This article provides a focused review of in situ hybridisation (ISH) methods.
The primary focus is on the detection of messenger RNA (mRNA) and micro RNA (miRNA).
Emphasis is placed on non-radioactive signal detection strategies applicable to formalin-fixed paraffin-embedded (FFPE) tissues.

Main Methods:

The review covers established ISH techniques adapted for FFPE samples.
It highlights strategies for detecting both mRNA and miRNA, which differ in size and processing.
A key aspect discussed is the implementation of non-radioactive detection systems for signal visualization.

Main Results:

Non-radioactive detection methods offer sensitive and specific visualization of mRNA and miRNA in FFPE tissues.
These methods provide direct spatial information on gene and microRNA expression within the tissue architecture.
The discussed strategies are routinely employed in laboratories for gene expression analysis.

Conclusions:

In situ hybridisation is a versatile and essential tool for visualizing nucleic acids in biological samples.
Non-radioactive ISH methods are highly effective for detecting mRNA and miRNA in FFPE tissues.
These techniques are crucial for understanding gene expression patterns and cellular function in various biological contexts.