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

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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.

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Data Availability

All data generated or analysed during this study are included in this published article and are available from the corresponding author on reasonable request.

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Acknowledgements

We acknowledge the aid provided by Prof. Aryeh Weiss in helpful discussions, to this work.

Author contributions

Concept and design: R.G., M.P. Data Acquisition: R.G., Z.G., and S.S. Data analysis: R.G., M.P., Z.G., and S.S. Data interpretation: R.G., M.P., Z.G., M.W., and I.L. Drafting the paper: R.G., M.P. Critical revision of the paper: All authors. Securing funding: R.G., E.M. Admin/technical/material support: R.G., E.M., I.L., and M.W. Supervision: R.G., E.M. Final approval: all authors.

Funding

Institute of Genetics—Soroka Medical Center.

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Author notes

  1. These authors contributed equally: Raphael Gonen, Max Platkov

Authors and Affiliations

  1. Nuclear Research Center Negev, 84190, Beer-Sheba, Israel

    Raphael Gonen, Max Platkov & Marcelo Weinstein

  2. Department of Biomedical Engineering, Ben Gurion University, Beer Sheva, Israel

    Raphael Gonen & Sheli Shayir

  3. Sami Shamoon College of Engineering, Beer‐Sheba, Israel

    Ziv Gardos & Inna Levitsky

  4. Department of Virology and Developmental Genetics, Faculty of Health Sciences, Ben Gurion University of the Negev, Beersheba, Israel

    Esther Manor

  5. Institute of Genetics, Soroka Medical Center and Faculty of Health Sciences, Ben Gurion University, Beer Sheba, Israel

    Esther Manor

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  1. Raphael Gonen
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Correspondence to Raphael Gonen.

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The authors declare no competing interests.

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Samples used in this study were obtained as part of routine medical care. Ethical approval for use of these samples for research purposes was not required for this study in accordance with local/national guidelines.

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Appendix

Appendix

This table shows the historical landmarks in the development of C-Banding techniques.

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Gonen, R., Platkov, M., Gardos, Z. et al. A DAPI-Based Modified C-banding Technique for a Rapid Achieving High Photographic Contrast of Centromeres on Chromosomes. Cell Biochem Biophys 80, 375–384 (2022). https://doi.org/10.1007/s12013-022-01065-5

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