Evaluation of manual and image analysis quantification of DNA damage in the alkaline comet assay

The alkaline comet assay or single cell microgel electrophoresis assay is a sensitive method of detecting DNA strand breaks and alkali labile sites in individual cells. The results of this assay can be analysed by different methods. In this study we compared analyses of the same slides by a manual m...

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Veröffentlicht in:Mutagenesis 1997-07, Vol.12 (4), p.209-214
Hauptverfasser: McCarthy, P.J., Sweetman, S.F., McKenna, P.G., McKelvey-Martin, V.J.
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container_end_page 214
container_issue 4
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container_title Mutagenesis
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creator McCarthy, P.J.
Sweetman, S.F.
McKenna, P.G.
McKelvey-Martin, V.J.
description The alkaline comet assay or single cell microgel electrophoresis assay is a sensitive method of detecting DNA strand breaks and alkali labile sites in individual cells. The results of this assay can be analysed by different methods. In this study we compared analyses of the same slides by a manual method and by image analysis, post-treatment of clone 707 Friend erythroleukaemia cells with H2O2. The parameters which were found to be particularly useful were comet area and comet length (measured manually) and percentage tail DNA, tail moment, tail length and tail length/head radius (L/H), measured using image analysis. The manual method for comet analysis presented in this paper would appear to provide good and reliable comet data. However, the image analysis comet system described offers an alternative analysis method which avoids the need for photomicrographs and tedious manual analysis. The image analysis parameters: % tail DNA, tail moment, tail length and L/H give good consistent results and for large-scale analysis it will, therefore, conceivably be the method of choice.
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source MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection
subjects Animals
Biological and medical sciences
Chemical mutagenesis
DNA - analysis
DNA - chemistry
DNA Damage - drug effects
Dose-Response Relationship, Drug
Electrophoresis, Agar Gel - methods
Hydrogen Peroxide - pharmacology
Image Processing, Computer-Assisted - methods
Leukemia, Erythroblastic, Acute - drug therapy
Leukemia, Erythroblastic, Acute - genetics
Medical sciences
Mice
Mutagenicity Tests
Oxidants - pharmacology
Reproducibility of Results
Toxicology
Tumor Cells, Cultured
title Evaluation of manual and image analysis quantification of DNA damage in the alkaline comet assay
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