Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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,...

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...

Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Transient and constitutive repression of cytoplasmic translation signaling in cells with mtDNA mutation.

Cellular and molecular life sciences : CMLS·2009

Same author

Overview of fluorescence in situ hybridization techniques for molecular cytogenetics.

Current protocols in cytometry·2008

Same author

Basic preparative techniques for fluorescence in situ hybridization.

Current protocols in cytometry·2008

Same author

Immunocytochemical detection.

Current protocols in cytometry·2008

Same author

COmbined Binary RAtio fluorescence in situ hybridiziation (COBRA-FISH): development and applications.

Cytogenetic and genome research·2006

Same author

Cryptic terminal chromosome rearrangements in colorectal carcinoma cell lines detected by subtelomeric FISH analysis.

Cytogenetic and genome research·2006

Search research articles

Related Experiment Video

Updated: Jul 2, 2026

Double Fluorescence in situ Hybridization in Fresh Brain Sections

Double Fluorescence in situ Hybridization in Fresh Brain Sections

Published on: August 14, 2010

Probe labeling and fluorescence in situ hybridization.

J Wiegant¹, A K Raap

¹Leiden University, Leiden, The Netherlands.

Current Protocols in Cytometry

|September 5, 2008

Summary

This study details essential protocols for in situ hybridization, including probe labeling, DNA denaturation, and washing steps. It also covers probe purification and quality control for accurate results.

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

In situ hybridization (ISH) is a powerful technique for visualizing nucleic acid targets within their cellular context.
Standardized and detailed protocols are crucial for the reproducibility and reliability of ISH experiments.

Purpose of the Study:

To provide a comprehensive guide to fundamental protocols for in situ hybridization.
To detail essential steps including probe labeling, target DNA denaturation, hybridization, and post-hybridization washes.

Main Methods:

Detailed description of probe labeling techniques, including purification and quality control.
Step-by-step instructions for denaturing in situ target DNA.
Protocols for performing in situ hybridization and subsequent post-hybridization washes.

More Related Videos

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

Published on: December 10, 2012

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Published on: March 25, 2009

Related Experiment Videos

Last Updated: Jul 2, 2026

Double Fluorescence in situ Hybridization in Fresh Brain Sections

Double Fluorescence in situ Hybridization in Fresh Brain Sections

Published on: August 14, 2010

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

Published on: December 10, 2012

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Published on: March 25, 2009

Main Results:

Establishment of robust and reproducible protocols for key steps in the in situ hybridization workflow.
Demonstration of effective probe purification and quality control methods to ensure hybridization efficiency.
Optimization of denaturation and washing steps for enhanced signal detection and reduced background.

Conclusions:

The described protocols provide a foundational framework for successful in situ hybridization experiments.
Adherence to these detailed methods ensures high-quality results for analyzing nucleic acid localization in biological samples.
This unit serves as a valuable resource for researchers utilizing in situ hybridization in various scientific disciplines.