Automatic analysis of slides processed in the Comet assay.
In recent years the Comet assay (or single cell gel electrophoresis assay) has been established as a rapid and sensitive method for the detection of DNA damage. For early genotoxicity screening of new chemical entities in industrial toxicology, the Comet assay is more and more used for assessment of the DNA damaging potential of a test compound. In order to increase compound screening throughput, we have established an image analysis system for fully automated measurement of microscope slides processed in the Comet assay. For the comparative investigation various cell types, such as V79 Chinese hamster cells, mouse lymphoma cells and human leukocytes, were treated with several test compounds. Using tail moment as the quantitative parameter for comet formation, we show a very high correlation between our automatic image analysis system and a commercially available, interactive system (Comet Assay II of Perceptive Instruments). The possibility of analyzing 50 samples within 1 day and the high reproducibility of results make automated image processing a powerful tool for automatic analysis of slides processed in the Comet assay.
Novartis Pharma AG, Preclinical Safety, Toxicology/Pathology, CH-4002 Basel, Switzerland. wilfried.frieauff@pharma.novartis.com
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D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D002478:Cells, Cultured; D020552:Comet Assay / Q000295:instrumentation* / Q000379:methods*; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D007940:Leukemia L5178; D008297:Male; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D009153:Mutagens / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar
D000873:Anthracenes; D009153:Mutagens; C006593:2-anthramine; D005020:Ethyl Methanesulfonate