title: DNA comet Giemsa staining for conventional bright-field microscopy.
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.
authors.affiliation: State Research Center-Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), 46, Zhivopisnaya Str., Moscow 123098, Russia. andreyan.osipov@gmail.com.
authors.forename: Andreyan
authors.initials: A
authors.lastname: Osipov
authors.affiliation: State Research Center-Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), 46, Zhivopisnaya Str., Moscow 123098, Russia. ek.arkhangelskaya@gmail.com.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Arkhangelskaya
authors.affiliation: Dima Rogachev Federal Research Center of Pediatric Hematology, Oncology and Immunology, 1, Samora Machel Str., Moscow 117997, Russia. a_vinokurov@inbox.ru.
authors.forename: Alexei
authors.initials: A
authors.lastname: Vinokurov
authors.affiliation: State Research Center-Burnasyan Federal Medical Biophysical Center of Federal Medical Biological Agency (SRC-FMBC), 46, Zhivopisnaya Str., Moscow 123098, Russia. smetaninanm@gmail.com.
authors.forename: Nadezhda
authors.initials: N
authors.lastname: Smetaninа
authors.affiliation: Dima Rogachev Federal Research Center of Pediatric Hematology, Oncology and Immunology, 1, Samora Machel Str., Moscow 117997, Russia. alex@biogerontology.org.
authors.forename: Alex
authors.initials: A
authors.lastname: Zhavoronkov
authors.affiliation: Chalk River Laboratories, Atomic Energy of Canada Limited, Stn 51 Plant Road, Chalk River, ON K0J1P0, Canada. klokovd@aecl.ca.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Klokov
mesh_terms: D000328:Adult; D052160:Benzothiazoles; D020552:Comet Assay; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage / Q000528:radiation effects; D003959:Diamines; D005260:Female; D006801:Humans; D008214:Lymphocytes / Q000473:pathology / Q000528:radiation effects; D008853:Microscopy; D009930:Organic Chemicals / Q000737:chemistry / Q000378:metabolism; D011804:Quinolines; D014965:X-Rays
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA
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