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 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.
McCarthy
P J
PJ
Cancer and Ageing Research Group, University of Ulster, Coleraine, N. Ireland, UK.
Sweetman
S F
SF
McKenna
P G
PG
McKelvey-Martin
V J
VJ
D000818:Animals; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D004915:Leukemia, Erythroblastic, Acute / Q000188:drug therapy / Q000235:genetics; D051379:Mice; D009152:Mutagenicity Tests; D016877:Oxidants / Q000494:pharmacology; D015203:Reproducibility of Results; D014407:Tumor Cells, Cultured
D016877:Oxidants; D004247:DNA; D006861:Hydrogen Peroxide