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Related Concept Videos

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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In-situ Hybridization02:31

In-situ Hybridization

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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...
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Double Fluorescence in situ Hybridization in Fresh Brain Sections
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Fluorescence In Situ Hybridization on Tissue Sections.

Alvin S T Lim1, Tse Hui Lim2

  • 1Cytogenetics Laboratory, Department of Molecular Pathology, Singapore General Hospital, Academia level 9, 20 College Road, Singapore, 169856, Singapore. alvin.lim.s.t@sgh.com.sg.

Methods in Molecular Biology (Clifton, N.J.)
|December 3, 2016
PubMed
Summary

This study details protocols for fluorescence in situ hybridization (FISH) on formalin-fixed paraffin-embedded (FFPE) tissues. The described methods enable genetic analysis of FFPE tumor specimens and cell blocks.

Keywords:
Fluorescence in situ hybridizationFormalin-fixed paraffin-embedded tissuesProbes

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Area of Science:

  • Molecular Biology
  • Genetics
  • Pathology

Background:

  • Formalin-fixed paraffin-embedded (FFPE) tissues are standard for analyzing solid tumors using fluorescence in situ hybridization (FISH).
  • Tissue microarrays and cell blocks from cell suspensions are also utilized for FISH analysis.
  • FISH assays detect gene copy number variations and gene fusions in tissue samples.

Purpose of the Study:

  • To describe laboratory protocols for interphase FISH analysis on FFPE tissue specimens.
  • To provide a comprehensive protocol applicable to various FFPE preparations.

Main Methods:

  • The protocol involves slide deparaffinization, pretreatment, and protease treatment.
  • Hybridization with specific nucleic acid probes follows, with subsequent washing and counterstaining.
  • The described methods are applicable to all FFPE preparations.

Main Results:

  • The described FISH protocol is effective for analyzing FFPE tissues, including tumor specimens.
  • The assay can be completed in 3 consecutive days, with options for a more rapid procedure.

Conclusions:

  • Standardized FISH protocols are crucial for accurate genetic analysis of FFPE tissues.
  • The presented methods facilitate the detection of genetic alterations in tumor samples using FFPE specimens.