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Updated: May 14, 2026
03:55
Robust Detection of Gene Amplification in Formalin-Fixed Paraffin-Embedded Samples by Fluorescence In Situ Hybridization
Published on: July 12, 2024
Quantitative fluorescence in situ hybridization on paraffin embedded tissue
Mario Ricciardi1, Mauro Krampera, Marco Chilosi
1Department of Pathology, University of Verona, Verona, Italy.
Methods in Molecular Biology (Clifton, N.J.)
|February 13, 2013
Summary
Quantitative fluorescence in situ hybridization (Q-FISH) is a method to measure telomere length in cells and tissues. This study details protocols for staining, acquiring, and analyzing human cells using Q-FISH for accurate telomere length evaluation.
Area of Science:
- Biotechnology
- Molecular Biology
- Genetics
Background:
- Telomere length is a critical factor in cellular aging and disease.
- Quantitative fluorescence in situ hybridization (Q-FISH) is an established method for telomere length assessment.
Purpose of the Study:
- To describe standardized protocols for Q-FISH.
- To enable accurate quantitative evaluation of telomere length in human cells and tissues.
Main Methods:
- Staining of cell preparations or human tissues with fluorescent peptide nucleic acid (PNA) probes targeting telomere sequences (5'-TTAGGG-3').
- Image acquisition using a fluorescence microscope with a sensitive CCD camera.
- Image analysis via specialized computer software for fluorescence intensity measurements.
Main Results:
- Detailed protocols for staining, image acquisition, and data analysis are presented.
- The described methods allow for quantitative measurement of telomere length.
Conclusions:
- The presented Q-FISH protocols provide a reliable framework for evaluating telomere length.
- Accurate telomere length measurement is crucial for research in aging and various diseases.
Related Concept Videos
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...
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,...