Video enhanced laser scanning microscopic analysis of DNA comet formation.
Laser Scanning Microscopy is a sensitive tool that provides a unique method of analyzing biological systems. Coupled with the Single Cell Gel assay, it allows for accurate and reproducible detection of DNA strand breaks. An understanding of the theory of DNA comet formation is lacking. Using dexamethasone induced apoptosis in murine thymocytes as a model for double strand breaks, we used video enhanced laser scanning microscopy to evaluate the leading edge of DNA migration in the single cell gel assay. In this system, comet length increases significantly within the first thirty seconds of electrophoresis, the greatest increase in length is completed within the first minute, and the first two minutes are important in significant increases in DNA migration during DNA comet formation.
Department of Microbiology, Brigham Young University, Provo, Utah 84602.
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Fairbairn
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Rowe
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O'Neill
D000818:Animals; D017209:Apoptosis; D004247:DNA / Q000032:analysis*; D003907:Dexamethasone / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D051379:Mice; D008853:Microscopy / Q000379:methods*; D013950:Thymus Gland / Q000166:cytology; D014741:Video Recording
D003907:Dexamethasone; D004247:DNA