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Open AccessCommunication
Int. J. Mol. Sci. 2014, 15(4), 6086-6095; doi:10.3390/ijms15046086

DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

1
State Research Center—Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), 46, Zhivopisnaya Str., Moscow 123098, Russia
2
Dima Rogachev Federal Research Center of Pediatric Hematology, Oncology and Immunology, 1, Samora Machel Str., Moscow 117997, Russia
3
Chalk River Laboratories, Atomic Energy of Canada Limited, Stn 51 Plant Road, Chalk River, ON K0J1P0, Canada
*
Author to whom correspondence should be addressed.
Received: 20 February 2014 / Revised: 18 March 2014 / Accepted: 26 March 2014 / Published: 10 April 2014
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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Abstract

This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. View Full-Text
Keywords: Giemsa stain; comet assay; bright-field microscopy; DNA damage Giemsa stain; comet assay; bright-field microscopy; DNA damage
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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Osipov, A.; Arkhangelskaya, E.; Vinokurov, A.; Smetaninа, N.; Zhavoronkov, A.; Klokov, D. DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy. Int. J. Mol. Sci. 2014, 15, 6086-6095.

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