Quantitative fluorescence in situ hybridization on paraffin embedded tissue

Methods Mol Biol. 2013;976:167-73. doi: 10.1007/978-1-62703-317-6_13.

Abstract

Quantitative fluorescence in situ hybridization (Q-FISH) is a complex technique for the quantitative evaluation of telomere length on cell preparations or on human tissues. The samples are stained with a fluorescent peptide nucleic acid (PNA) probe against the telomere oligonucleotides (sequence 5'-TTAGGG-3'). The measure of the telomere length is carried out using a fluorescence microscope equipped with a sensitive CCD camera and analyzing the pictures with a computer software that can perform fluorescence intensity measurements. Here, we describe the most used protocols to stain, acquire, and analyze fixed human cells in order to evaluate their telomere length.

MeSH terms

  • Flow Cytometry
  • Humans
  • Image Processing, Computer-Assisted
  • In Situ Hybridization, Fluorescence / methods*
  • Microscopy, Fluorescence / methods*
  • Paraffin Embedding / methods*
  • Peptide Nucleic Acids / analysis*
  • Telomere / genetics

Substances

  • Peptide Nucleic Acids