Quantitative Fluorescence In Situ Hybridization (QFISH)
Fluorescence in situ hybridization (FISH) has a wide spectrum of applications in current molecular cytogenetic and cancer research. This is a unique technique that can be used for chromosomal DNA analysis in all cell types, at all stages of the cell cycle, and at molecular resolution. Recent developments in microscopy and imaging systems have allowed quantification of digital FISH images (quantitative FISH or QFISH) and have provided a new way for molecular cytogenetic analysis at single-cell level. QFISH can be applied for studying chromosome imbalances in interphase nuclei or metaphase spreads, measuring relative DNA content at chromosomal loci and identifying parental origin of homologous chromosomes. Here, a QFISH protocol suitable for the majority of DNA probes using the popular US National Institute of Health developed ImageJ software is described.
Key wordsChromosome abnormalities DNA probes Fluorescence in situ hybridization Interphase Nucleus Quantification QFISH
The article is dedicated to Ilia V. Soloviev. I would like to express my gratitude to Prof. Svetlana G Vorsanova and Prof. Yuri B Yurov for helping in the preparation of this chapter. This work was supported by the Russian Science Foundation (Grant #14-35-00060).
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