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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,...

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Robust 3D DNA FISH Using Directly Labeled Probes

Robust 3D DNA FISH Using Directly Labeled Probes

Published on: August 15, 2013

Combination DNA/RNA FISH and immunophenotyping.

¹Leiden University Medical Centre, Leiden, The Netherlands.

Current Protocols in Cytometry

|September 5, 2008

Summary

This summary is machine-generated.

This study details methods for combining immunophenotyping with DNA/RNA FISH for genotype/phenotype analysis. These protocols help identify chromosomal aberrations in heterogeneous cell populations by detecting DNA, RNA, and proteins simultaneously.

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Area of Science:

Cell Biology
Genetics
Molecular Biology

Background:

Analyzing heterogeneous cell populations requires methods to identify specific subpopulations.
Simultaneous detection of genetic material (DNA/RNA) and protein expression is challenging.
Chromosomal aberrations can be indicative of various diseases, necessitating precise detection methods.

Purpose of the Study:

To present protocols for integrating immunophenotyping with DNA/RNA FISH.
To enable genotype/phenotype analysis in complex cellular samples.
To facilitate the identification of chromosomal aberrations in specific cell subpopulations.

Main Methods:

Combining immunophenotyping (protein detection) with Fluorescence In Situ Hybridization (FISH) for DNA and RNA.
Developing protocols for simultaneous detection of nucleic acids and proteins within cells.
Applying these methods to heterogeneous cell populations for subpopulation analysis.

Main Results:

Successful integration of immunophenotyping with DNA/RNA FISH techniques.
Demonstration of genotype/phenotype analysis capabilities.
Identification of chromosomal aberrations within distinct cell subpopulations.

Conclusions:

The presented protocols enable robust combination of immunophenotyping and FISH.
This integrated approach is effective for analyzing chromosomal aberrations in heterogeneous cell samples.
The methods facilitate detailed genotype/phenotype characterization of cellular subpopulations.