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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...

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Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections

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Basic preparative techniques for fluorescence in situ hybridization.

J Wiegant¹, A K Raap

¹Leiden University, Leiden, The Netherlands.

Current Protocols in Cytometry

|September 5, 2008

Summary

This summary is machine-generated.

This study details protocols for preparing human chromosome slides and nuclei for Fluorescence In Situ Hybridization (FISH). These methods enable molecular cytogenetics analysis across various sample types.

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Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Fluorescence In Situ Hybridization (FISH) is a powerful molecular cytogenetics technique.
Effective FISH analysis relies on high-quality sample preparation.
Diverse biological samples require tailored preparation methods.

Purpose of the Study:

To present standardized protocols for preparing human chromosome and nuclear samples.
To ensure sample preparations are suitable for FISH analysis.
To provide methods applicable to various sample types, including blood and tissue.

Main Methods:

Protocols for preparing human metaphase chromosome slides from peripheral blood lymphocytes.
Methods for isolating interphase nuclei from lymphocytes and paraffin-embedded tissues.
Techniques for preparing DNA fibers for molecular analysis.

Main Results:

The presented protocols yield preparations amenable to FISH.
The methods cover a range of sample types relevant to FISH applications.
Successful application depends on the specific molecular cytogenetics question.

Conclusions:

Standardized protocols are crucial for reliable FISH-based molecular cytogenetics.
The choice of sample preparation method is critical and depends on the research question.
These protocols facilitate advanced genetic and cellular analysis using FISH.