Literature DB >> 35137344

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

Raphael Gonen1,2, Max Platkov3, Ziv Gardos4, Sheli Shayir5, Inna Levitsky4, Marcelo Weinstein3, Esther Manor6,7.   

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.
© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

Entities:  

Keywords:  C-Banding; Centromere Highlighting; Chromosome Highlighting; DAPI

Mesh:

Substances:

Year:  2022        PMID: 35137344     DOI: 10.1007/s12013-022-01065-5

Source DB:  PubMed          Journal:  Cell Biochem Biophys        ISSN: 1085-9195            Impact factor:   2.989


  13 in total

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2.  New tool for biological dosimetry: reevaluation and automation of the gold standard method following telomere and centromere staining.

Authors:  Radhia M'kacher; Elie E L Maalouf; Michelle Ricoul; Leonhard Heidingsfelder; Eric Laplagne; Corina Cuceu; William M Hempel; Bruno Colicchio; Alain Dieterlen; Laure Sabatier
Journal:  Mutat Res       Date:  2014-09-28       Impact factor: 2.433

3.  Automating dicentric chromosome detection from cytogenetic biodosimetry data.

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Journal:  Radiat Prot Dosimetry       Date:  2014-04-21       Impact factor: 0.972

4.  Automated discrimination of dicentric and monocentric chromosomes by machine learning-based image processing.

Authors:  Yanxin Li; Joan H Knoll; Ruth C Wilkins; Farrah N Flegal; Peter K Rogan
Journal:  Microsc Res Tech       Date:  2016-03-01       Impact factor: 2.769

5.  New C-band protocol by heat denaturation in the presence of formamide.

Authors:  R Fernández; M J L Barragán; M Bullejos; J A Marchal; R Díaz De La Guardia; A Sanchez
Journal:  Hereditas       Date:  2002       Impact factor: 3.271

6.  Polymorphism of human C-band heterochromatin. I. Frequency of variants.

Authors:  A P Craig-Holmes; F B Moore; M W Shaw
Journal:  Am J Hum Genet       Date:  1973-03       Impact factor: 11.025

7.  A simple technique for demonstrating centromeric heterochromatin.

Authors:  A T Sumner
Journal:  Exp Cell Res       Date:  1972-11       Impact factor: 3.905

8.  DNA sequence dependent binding modes of 4',6-diamidino-2-phenylindole (DAPI).

Authors:  W D Wilson; F A Tanious; H J Barton; R L Jones; K Fox; R L Wydra; L Strekowski
Journal:  Biochemistry       Date:  1990-09-11       Impact factor: 3.162

9.  Biological estimates of dose to inhabitants of Belarus and Ukraine following the Chernobyl accident.

Authors:  A Edwards; P Voisin; I Sorokine-Durm; N Maznik; V Vinnikov; L Mikhalevich; J Moquet; D Lloyd; M Delbos; V Durand
Journal:  Radiat Prot Dosimetry       Date:  2004-07-20       Impact factor: 0.972

10.  Cytogenetic biodosimetry for Fukushima travelers after the nuclear power plant accident: no evidence of enhanced yield of dicentrics.

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  1 in total

1.  Metaphase Cells Enrichment for Efficient Use in the Dicentric Chromosome Assay.

Authors:  Max Platkov; Uzi Hadad; Ariela Burg; Inna Levitsky; Michael Zagatzki; Omer Damri; Aryeh Weiss; Yair Lauber; Shirly Amar; Lior Carmel; Raphael Gonen
Journal:  Cell Biochem Biophys       Date:  2022-10-11       Impact factor: 2.989

  1 in total

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