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.

[1]  A. Wyllie,et al.  Apoptosis. The role of the endonuclease. , 1990, The American journal of pathology.

[2]  A. Whetton,et al.  Induction of apoptosis--new targets for cancer chemotherapy. , 1992, Seminars in cancer biology.

[3]  K. O’Neill,et al.  Application of confocal laser scanning microscopy to analysis of H2O2-induced DNA damage in human cells. , 1993, Scanning.

[4]  P. Olive,et al.  DNA double-strand breaks measured in individual cells subjected to gel electrophoresis. , 1991, Cancer research.

[5]  K J Johanson,et al.  Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. , 1984, Biochemical and biophysical research communications.

[6]  A. Collins,et al.  Single-cell gel electrophoresis applied to the analysis of UV-C damage and its repair in human cells. , 1992, International journal of radiation biology.

[7]  I. Olivotto,et al.  Gel electrophoresis of individual cells to quantify hypoxic fraction in human breast cancers. , 1993, Cancer research.

[8]  R E Durand,et al.  Factors influencing DNA migration from individual cells subjected to gel electrophoresis. , 1992, Experimental cell research.

[9]  Vijayalaxmi,et al.  Assessment of radiation-induced DNA damage in human blood lymphocytes using the single-cell gel electrophoresis technique. , 1992, Mutation research.

[10]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[11]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.