Microscopy-based approaches to the use of radiation-induced nuclear foci as biomarkers of DNA repair.
Published: 2006
Language: eng
Number of pages: 155
ISBN-13: 9780494212615
Description: The visualization of ionizing radiation-induced foci (IRIF) by immunofluorescent microscopy is a proven method of identifying sites of DNA double strand breaks (DNA-DSB). IRIF can be analysed using high-resolution microscopes with appropriate filter sets. In this thesis, I developed a rigorous microscopic approach to IRIF foci image analysis with the development of appropriate algorithms. I then tested the hypothesis that H2AXS139ph IRIF are dose and time responsive and can reflect differences in DNA-DSB repair using normal and DNA-DSB repair-deficient human fibroblasts. The algorithms for automatic foci counting utilized wide-field microscopic deconvolution and multi-pass filtering to determine "true" IRIF over background. Initial H2AXS139ph IRIF can be quantitated using this technique, but it was found that H2AXS139ph IRIF did not predict DNA-DSB repair between the two cell types. Using the methods developed in this thesis, investigators can now pursue dedicated DNA-dsb experiments with an improved "signal to noise" ratio.
Notes: Source: Masters Abstracts International, Volume: 45-03, page: 1413.

Thesis (M.Sc.)--University of Toronto, 2006.

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