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

The alkaline comet assay or single cell microgel electrophoresisassay is a sensitive method of detecting DNA strand breaks andalkali labile sites in individual cells. The results of thisassay can be analysed by different methods. In this study wecompared analyses of the same slides by a manual method andby image analysis, post-treatment of clone 707 Friend erythroleukaemiacells with H₂O₂. The parameters which were found to be particularlyuseful were comet area and comet length (measured manually)and percentage tail DNA, tail moment, tail length and tail length/headradius (L/H), measured using image analysis. The manual methodfor comet analysis presented in this paper would appear to providegood and reliable comet data. However, the image analysis cometsystem described offers an alternative analysis method whichavoids the need for photomicrographs and tedious manual analysis.The image analysis parameters: % tail DNA, tail moment, taillength and L/H give good consistent results and for large-scaleanalysis it will, therefore, conceivably be the method of choice. ¹To whom correspondence should be addressed