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Micro-irradiation systems are used to analyse the effect of ionizing radiation at the cellular and tissue level, targeting individual cells within a population with a controlled low dose. Cell survival experiments using micro-irradiation systems are limited by factors including: 1) the radiation attenuation and optical properties of the chosen cell dish substrate, 2) the registration of the cell dish before and after irradiation or between multiple imaging modalities and 3) the analysis of the cell or colony growth after irradiation. In this thesis, a set of tools have been developed to improve micro-irradiation experiments and to increase the accuracy of information provided by the cell survival data. The first contribution, the substrate cell dish evaluation, provides a set of characteristics defining the substrates used for micro-irradiation experiments based on minimal energy loss and optical clarity using unstained cell imaging. The second contribution was the development of a novel and low cost fiducial marking device for micro-irradiation experiments using an 808 nm laser and providing marks suitable for imaging with multiple modalities. The minimum focused spot diameter was calculated as 22.9 urn and the device was used to create fiducial marks with diameters ranging from 20 urn to 130 urn. The third contribution, the development of a cell counting methodology for use with a lens-free imaging device, has been shown to accurately count thousands of cells suitable for immediate analysis. Approximately 1000 cell colonies, containing 17 729 cells on 11 cell dishes were used for testing and training for automatic cell counting. Validation of the cell counting method showed that 76% and 89% of the cell colonies were counted within a ± 20% and ± 30% error of the ground truth, respectively. Further development of the fiducial marking device, by modifying the choice of laser and making it suitable for multiple types of cell dish substrates, would increase the applications of the device. Development of the cell counting methodology for different cells line, and for cells grown on multiple types of substrates, would make the system suitable for analysis of a wide variety of cell survival studies. The cell counting methodology, applied to the CyMap lens-free imaging device, has the potential to be an extremely useful and cost effective tool for cell survival studies.

Thesis (DTech (Biomedical Technology))--Cape Technikon, 2001 / Ionising radiation has the ability to induce, inter alia, DNA damage and is well established as a causative agent of carcinogenesis and mutagenesis. The effects of high doses of short duration have been well documented, whereas the effects of continuous exposure to low doses of ionising radiation have not, nor are they as clearly understood and current risk estimates are largely extrapolated from high-dose data of atomic bomb survivors. This study evaluated the clastogenic effects of low dose ionising radiation on a population of bats (Chiroptera) residing in an abandoned monazite mine. Bats were sampled from two areas in the mine, with external radiation levels measuring around 20 µSv/h (low dose) and 100 µSv/h (high dose). A control group of bats was collected from a cave with no detectable radiation above normal background levels. The most frequently encountered genetic event in human malignancy is the alteration of the p53 gene. Mutant p53 proteins have a longer half-life than the wild-type variant and accumulate to high levels in the nucleus of tumour cells. The study showed that not only was there a significant increase in p53 positive cells of radiation exposed bats, but also in the degree of positivity, especially in the cells lining the bronchioles of the lungs. Although experimental studies have shown that exposure to radiation may lead to the onset of fibrosis and an inflammatory response in the lung and other tissues, the magnitude of the dose exposure was not comparable to this study and histological examination of bat lung and liver tissues showed no morphological changes in radiation exposed bats when compared to the control group. It has been documented that chronic radiation exposures may give rise to a number of specific haematological defects which are collectively termed "preleukemia" or myelodysplastic syndrome. Full blood counts on bat samples showed a significant decrease in the MCV indicating microcytic erythrocytes from the radiation exposed bats. Differential counts performed on the peripheral blood of the bats showed a marked neutropenia. Neutrophils also showed marked dysplasia including psuedo-Pelger Huet cells in radiation-exposed bats. Cytochemical analysis using DAB myeIoperoxidase showed that control bats had hypogranular neutrophils andradiation-exposedbats had largely '1granularneutrophils. Bonemarrow biopsies were taken from both control and radiation-exposed bats and evaluated for ceIlularity, granulocyte: lymphocyte: erythrocyte (GLE) ratio and megakaryocyte morphology. A hypocelIular bone marrow, a decreased granulocytic haematopoeisis and dysplastic megakaryocyte morphology were observed in radiation-exposed bats. Mineralisation of bone osteoid was determined using image analysis and showed a highly significant decrease in the bone matrix from radiation-exposed bats. All haematological features observed are congruent with current literature describing secondary (radiation-induced) myelodysplastic syndrome.

Thesis (M.S.)--Uniformed Services University of the Health Sciences, 2005. / Typescript (photocopy).