Application of confocal laser scanning microscopy to analysis of H2O2‐induced DNA damage in human cells
Open Access
- 1 January 1993
- Vol. 15 (3) , 136-139
- https://doi.org/10.1002/sca.4950150305
Abstract
Confocal laser scanning microscopy (CLSM) offers improved depth discrimination and spatial resolution to the analysis of biologic samples. We demonstrate in this paper that such technology is valuable in examining DNA single‐strand breaks in human cells. The single‐cell‐gel (SCG) assay is a new technique for measuring DNA strand breaks in individual cells. Cells embedded in lowmelting‐point agarose are treated with varying concentrations of hydrogen peroxide to induce DNA strand breaks. Following cell lysis and alkaline electrophoresis, which enables single‐stranded break detection, analysis of the resulting “comets” provides an accurate method of comparing changes in DNA migration patterns, which have been shown to reflect the DNA damage levels. A statistically significant difference (p < 0.01) in single‐stranded DNA damage levels was detected in cells exposed to hydrogen peroxide concentrations as low as 10 nm for 2 min. LSM analysis of the SCG technique allows rapid, sensitive and reproducible quantitation of single‐stranded breaks of cellular DNA.Keywords
This publication has 12 references indexed in Scilit:
- Colour thresholding and objective quantification in bioimagingJournal of Microscopy, 1992
- Chromosomal localization of transfected genes by a combination of hot banding and fluorescence in situ hybridization.Journal of Histochemistry & Cytochemistry, 1992
- Assessment of radiation-induced DNA damage in human blood lymphocytes using the single-cell gel electrophoresis techniqueMutation Research/Environmental Mutagenesis and Related Subjects, 1992
- Factors influencing DNA migration from individual cells subjected to gel electrophoresisExperimental Cell Research, 1992
- Induction and Rejoining of DNA Single-Strand Breaks in Relation to Cellular Growth in Human Cells Exposed to Three Hydroperoxides at 0°C and 37°CFree Radical Research Communications, 1991
- A simple technique for quantitation of low levels of DNA damage in individual cellsExperimental Cell Research, 1988
- An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy.The Journal of cell biology, 1987
- Prooxidant States and Tumor PromotionScience, 1985
- Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cellsBiochemical and Biophysical Research Communications, 1984