A DAPI-Based Modified C-banding Technique for a Rapid Achieving High Photographic Contrast of Centromeres on Chromosomes

Raphael Gonen, Max Platkov, Ziv Gardos, Sheli Shayir, Inna Levitsky, Marcelo Weinstein, Esther Manor

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Many chromosome assays rely on the quantification of chromosome abnormalities in cells, and one important abnormality is the existence of more than one centromere for each chromosome. The quantification of such abnormalities has been studied before. However, this process is labor-intensive and time consuming. Thus, this assay is challenging for ex-laboratory applications, where speed is required. We present a visualization method that uses a cheap stain—DAPI, long (e.g., high-resolution) chromosomes and our modified C-banding method for labeling chromosomes. The labeled chromosomes can then be easily seen with a conventional and readily available fluorescence microscopy system. This method achieves an acceleration of the detection of the presence of constitutive heterochromatin in chromosomal centromeres by more than 10 times, to ~2 h, in Human lymphocyte cells and in cells of the human Jurkat line. This new procedure will ultimately provide an easier and cheaper alternative to FISH/PNA probes, or the classic Giemsa staining method. Simplification and reduction in time of the overall procedure will enable the utilization of centromere-counting assays in laboratory and ex-laboratory applications, including in emergency response.

Original languageEnglish
Pages (from-to)375-384
Number of pages10
JournalCell Biochemistry and Biophysics
Volume80
Issue number2
DOIs
StatePublished - 1 Jun 2022

Keywords

  • C-Banding
  • Centromere Highlighting
  • Chromosome Highlighting
  • DAPI

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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