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FISH is a well-established technique, which was developed in the early 1980s [1-4]. It is often performed as a single gene, break-apart assay that detects DNA level rearrangements.

The basic steps in a FISH protocol include:

  1. Use of fluorescent nucleic acid probes to detect and bind to specific DNA sequences in intact cells or tissue (commercial probes for ETV6, NTRK1, and NTRK3 are available, as well as fusions probes for combined ETV6-NTRK3 detection).
  2. Fluorescence microscopy visualisation of the fluorescent probe once it is bound to a chromosome or RNA.


  1. Savic S, Bubendorf L. Common Fluorescence In Situ Hybridization Applications in Cytology. Arch Pathol Lab Med 2016; 140: 1323-1330.
  2. Langer-Safer PR, Levine M, Ward DC. Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A 1982; 79: 4381-4385.
  3. Chae YK, Arya A, Chiec L et al. Challenges and future of biomarker tests in the era of precision oncology: Can we rely on immunohistochemistry (IHC) or fluorescence in situ hybridization (FISH) to select the optimal patients for matched therapy? Oncotarget 2017; 8: 100863-100898.
  4. Urbanek MO, Nawrocka AU, Krzyzosiak WJ. Small RNA Detection by in Situ Hybridization Methods. Int J Mol Sci 2015; 16: 13259-13286.

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