A Quantitative Analysis of Radiation-Induced Chromosome Aberrations with a Fluorescent Digital Image Microscope

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Using a synthetic, alpha satellite consensus DNA unit as a probe, we could show dicentrics as well as acentric fragmented chromosomes in metaphases of γ-irradiated lymphocyte cells and that the number of appearance of dicentrics or acentric fragments seemed to be proportional to radiation doses. To make such examination with a large number of chromosomes of metaphases, a quantitative fluorescence measurement was performed using a fluorescent microscope digital image analysis system. The relative amounts of fluoresceinated probe hybridized to alpha satellite DNA varied with chromo-somes to a certain extent (1-4% of total probe-fluorescence of one metaphase). However, we could score dicentrics and acentric fragments as dots with extraordinarily higher or low percentage in plots of relative amount of probe-fluorescence on metaphase chromosome in γ-irradiated cells. The number of appearacne of acentric fragments were proportional to radiation doses of 1, 2 and 4 Gy. In the case of dicentrics, the number of appearance seemed to be proportional to square doses. Further, to reduce the variation of relative amounts of probe-fluorescence, we tried to make probe-fluorescence reflect the content of alpha satellite DNA on each chromosome more exactly, using a DNA probe amplified by polymerase chain reaction.
日本放射線影響学会の論文