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Mutagenesis Advance Access published online on January 12, 2009
Mutagenesis, doi:10.1093/mutage/gen071
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© The Author 2009. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org.

A direct view by immunofluorescent comet assay (IFCA) of DNA damage induced by nicking and cutting enzymes, ionizing 137Cs radiation, UV-A laser microbeam irradiation and the radiomimetic drug bleomycin

Paulius Grigaravicius1,*, Alexander Rapp1,2 and Karl Otto Greulich1

1Leibniz Institute for Age Research—Fritz Lipmann Institute, Beutenbergstrasse 11, D-07745 Jena, Germany

In DNA repair research, DNA damage is induced by different agents, depending on the technical facilities of the investigating researchers. A quantitative comparison of different investigations is therefore often difficult. By using a modified variant of the neutral comet assay, where the histone H1 is detected by immunofluorescence [immunofluorescent comet assay (IFCA)], we achieve previously unprecedented resolution in the detection of fragmented chromatin and show that trillions of ultraviolet A photons (of a few eV), billions of bleomycin (BLM) molecules and thousands of {gamma} quanta (of 662 keV) generate, in first order, similar damage in the chromatin of HeLa cells. A somewhat more detailed inspection shows that the damage caused by 20 Gy ionizing radiation and by a single laser pulse of 10 µJ are comparable, while the damage caused by 12 µg/ml BLM depends highly on the individual cell. Taken together, this work provides a detailed view of DNA fragmentation induced by different treatments and allows comparing them to some extent, especially with respect to the neutral comet assay.

* To whom correspondence should be addressed. Tel: +49 3641 656409; Fax: +49 3641 656410; Email: paulius{at}fli-leibniz.de

2 Present address: Department of Biology, Technische Universitaet Darmstadt, Schnittspahnstrasse 10, D-64287 Darmstadt, Germany

Received on August 4, 2008; revised on November 14, 2008; accepted on November 16, 2008.


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