Detection and observation of oxidative damage for the improvement of high-LET radiation therapy

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In order to visualize the distribution of a penumbra, a low LET region around ion tracks, we have developed the methods to image the distribution of 8-OHdG, a radiation product resulting from the reaction of OH radicals with guanine residue, using an immunofluorescence staining for 1) mouse brain tissue, 2) human A549 cells, and 3) DNA thin film. Positive stains were evident particularly in the peripheral region of the cerebrum and in Purkinje cells of the cerebellum in mice. The immunofluorescence staining of cell nuclei was detected at a dose as small as 5 Gy where we can distinguish each incoming particle. DNA film with an insoluble property to water was developed with special reference to the uniformity, and the treatment with Fenton reaction to generate OH radicals resulted in the successful staining of uniformly formed 8-OHdG.