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07:36
Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections
Published on: June 25, 2021
Fluorescence in situ hybridization methods and troubleshooting applied to fixed cell suspensions
John Swansbury1, Toon Min, Shilani Aruliah
1Clinical Cytogenetics Laboratory, The Royal Marsden Hospital and The Institute of Cancer Research, Sutton, Surrey, UK. john.swansbury@rmh.nhs.uk
Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
Summary
This study details a reliable fluorescence in situ hybridization (FISH) procedure, offering troubleshooting advice. Enhanced probe availability makes FISH a crucial tool for cytogeneticists studying chromosome abnormalities.
Area of Science:
- Molecular Biology
- Genetics
- Cytogenetics
Background:
- Fluorescence in situ hybridization (FISH) is a key cytogenetic technique.
- Advances in FISH probe reliability and commercial availability have increased its utility.
- Understanding chromosome abnormalities is crucial for interpreting FISH results.
Purpose of the Study:
- To describe a robust and reliable fluorescence in situ hybridization (FISH) procedure.
- To provide practical troubleshooting tips for FISH assays.
- To highlight the importance of FISH as a complementary tool in cytogenetics.
Main Methods:
- Detailed protocol for performing fluorescence in situ hybridization (FISH).
- Inclusion of troubleshooting guidance for common FISH experimental issues.
- Integration of FISH with karyotype analysis.
Main Results:
- A validated and robust FISH procedure is presented.
- Effective troubleshooting strategies are provided to ensure reliable results.
- Demonstration of FISH as an essential adjunct to karyotyping.
Conclusions:
- The described FISH procedure is reliable and essential for cytogeneticists.
- FISH is a powerful tool for identifying chromosome abnormalities.
- Knowledge of chromosome abnormalities enhances the interpretation of FISH signals.
Related Concept Videos
02:07FISH - 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,...
02:31In-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...
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...