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dc.creatorBulat, Tanja M.
dc.creatorKeta, Otilija D.
dc.creatorKorićanac, Lela
dc.creatorŽakula, Jelena
dc.creatorPetrović, Ivan M.
dc.creatorRistić-Fira, Aleksandra
dc.creatorTodorović, Danijela V.
dc.date.accessioned2018-03-01T16:44:43Z
dc.date.available2018-03-01T16:44:43Z
dc.date.issued2016
dc.identifier.issn0001-3765 (print)
dc.identifier.issn1678-2690 (electronic)
dc.identifier.urihttp://vinar.vin.bg.ac.rs/handle/123456789/970
dc.description.abstractIonizing radiation induces DNA double strand breaks (DSBs) that trigger phosphorylation of the histone protein H2AX (gamma H2AX). Immunofluorescent staining visualizes formation of gamma H2AX foci, allowing their quantification. This method, as opposed to Western blot assay and Flow cytometry, provides more accurate analysis, by showing exact position and intensity of fluorescent signal in each single cell. In practice there are problems in quantification of gamma H2AX. This paper is based on two issues: the determination of which technique should be applied concerning the radiation dose, and how to analyze fluorescent microscopy images obtained by different microscopes. HTB140 melanoma cells were exposed to gamma-rays, in the dose range from 1 to 16 Gy. Radiation effects on the DNA level were analyzed at different time intervals after irradiation by Western blot analysis and immunofluorescence microscopy. Immunochemically stained cells were visualized with two types of microscopes: AxioVision (Zeiss, Germany) microscope, comprising an ApoTome software, and AxioImagerA1 microscope (Zeiss, Germany). Obtained results show that the level of gamma H2AX is time and dose dependent. Immunofluorescence microscopy provided better detection of DSBs for lower irradiation doses, while Western blot analysis was more reliable for higher irradiation doses. AxioVision microscope containing ApoTome software was more suitable for the detection of gamma H2AX foci.en
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/173046/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/171019/RS//
dc.rightsopenAccessen
dc.sourceAnais de Academia Brasileira de Cienciasen
dc.subjectApoTome softwareen
dc.subjectAxioImagerA1 microscopeen
dc.subjectimmunofluorescence microscopyen
dc.subjectWestern bloten
dc.subjectgamma H2AXen
dc.titleRadiation dose determines the method for quantification of DNA double strand breaksen
dc.typearticleen
dcterms.abstractКета, Отилија; Булат Тања М.; Тодоровиц, Данијела; Корићанац Лела; Жакула Јелена; Петровић Иван; Ристић-Фира Aлександра;
dc.citation.volume88
dc.citation.issue1
dc.citation.spage127
dc.citation.epage136
dc.identifier.wos000372172900011
dc.identifier.doi10.1590/0001-3765201620140553
dc.citation.rankM23
dc.identifier.pmid26959322
dc.identifier.scopus2-s2.0-84960461391
dc.identifier.fulltexthttp://vinar.vin.bg.ac.rs//bitstream/id/14345/966.pdf


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