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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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Quantitative Fluorescence In Situ Hybridization (QFISH).

Ivan Y Iourov1,2,3

  • 1Mental Health Research Center, Zagorodnoe sh. 2, Moscow, 117152, Russia. ivan.iourov@gmail.com.

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

Quantitative FISH (qFISH) offers precise DNA analysis at the single-cell level for cancer research. This study details a qFISH protocol using ImageJ software for broad DNA probe compatibility in molecular cytogenetics.

Keywords:
Chromosome abnormalitiesDNA probesFluorescence in situ hybridizationInterphaseNucleusQFISHQuantification

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Area of Science:

  • Molecular Cytogenetics
  • Cancer Research
  • Biotechnology

Background:

  • Fluorescence in situ hybridization (FISH) is a key technique for chromosomal DNA analysis.
  • FISH provides molecular resolution across all cell types and cell cycle stages.
  • Advancements in imaging enable digital FISH image quantification (qFISH) for single-cell analysis.

Purpose of the Study:

  • To describe a quantitative FISH (qFISH) protocol.
  • To ensure broad DNA probe compatibility.
  • To utilize the ImageJ software for molecular cytogenetic analysis.

Main Methods:

  • Development of a qFISH protocol.
  • Application of the protocol with various DNA probes.
  • Utilizing ImageJ software for image analysis and quantification.

Main Results:

  • The described qFISH protocol is suitable for most DNA probes.
  • The protocol facilitates molecular cytogenetic analysis at the single-cell level.
  • ImageJ software enables accurate quantification of digital FISH images.

Conclusions:

  • Quantitative FISH (qFISH) provides a powerful method for molecular cytogenetic analysis.
  • The described ImageJ-based qFISH protocol enhances accessibility and application in research.
  • qFISH is valuable for studying chromosome imbalances, DNA content, and chromosomal abnormalities.