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FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

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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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Related Experiment Video

Updated: Jun 11, 2025

Modified Terminal Restriction Fragment Analysis for Quantifying Telomere Length Using In-gel Hybridization
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Modified Terminal Restriction Fragment Analysis for Quantifying Telomere Length Using In-gel Hybridization

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Telomere Length Measurement in Human Tissue Sections by Quantitative Fluorescence In Situ Hybridization (Q-FISH).

Keisuke Nonaka1, Junko Aida2, Yasuko Hasegawa2

  • 1Research Team for Geriatric Pathology, Tokyo Metropolitan Institute for Geriatrics and Gerontology, Tokyo, Japan. nona_kei@tmig.or.jp.

Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2024
PubMed
Summary

Telomere shortening is a key feature of cellular senescence. This study details a quantitative fluorescence in situ hybridization (Q-FISH) method to measure relative telomere lengths in human tissue sections.

Keywords:
CentromereFFPEFormalin-fixed paraffin-embeddedHuman tissuePNA probeQ-FISHQuantitative fluorescence in situ hybridizationRelative telomere lengthTelomere

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Area of Science:

  • Cell Biology
  • Genetics
  • Histology

Background:

  • Telomeres shorten with cell division, leading to cellular senescence.
  • Telomere shortening is a hallmark of aging and disease.
  • Assessing telomere length in human tissues is crucial for understanding cellular aging.

Purpose of the Study:

  • To describe quantitative fluorescence in situ hybridization (Q-FISH) protocols.
  • To enable the measurement of relative telomere lengths in formalin-fixed paraffin-embedded (FFPE) human tissue sections.
  • To provide a method for assessing cellular senescence in histological samples.

Main Methods:

  • Utilizing quantitative fluorescence in situ hybridization (Q-FISH).
  • Employing fluorescently labeled peptide nucleic acid (PNA) probes for telomeric and centromeric sequences.
  • Hybridizing probes to FFPE human tissue sections.
  • Measuring relative telomere lengths by comparing telomere and centromere signal intensities.

Main Results:

  • The Q-FISH method allows for the estimation of telomere lengths in individual cells.
  • Relative telomere lengths can be accurately measured in FFPE human tissue sections.
  • The protocols described enable robust assessment of telomere status in diverse tissue types.

Conclusions:

  • Q-FISH is a valuable technique for studying telomere dynamics in human tissues.
  • This method facilitates research into the role of telomere shortening in cellular senescence and disease.
  • The described protocols provide a standardized approach for telomere length analysis in FFPE samples.