DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

International Journal of Molecular Sciences

Andreyan Osipov²  Ekaterina Arkhangelskaya²  Alexei Vinokurov¹  Nadezhda Smetanina²  Alex Zhavoronkov¹ 

[1] Dima Rogachev Federal Research Center of Pediatric Hematology, Oncology and Immunology, 1, Samora Machel Str., Moscow 117997, Russia; E-Mails:;State Research Center—Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), 46, Zhivopisnaya Str., Moscow 123098, Russia; E-Mails:

关键词: Giemsa stain;    comet assay;    bright-field microscopy;    DNA damage;   

DOI  :  10.3390/ijms15046086

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland