Detection and evaluation of chromosomal aberrations induced by high doses of-irradiation using immunogold-silver painting of prematurely condensed chromosomes

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Abstract
We developed a method for detecting chromosomal aberrations after irradiation with high doses of gamma-rays. At high dose levels, few of the irradiated cells are able to enter mitosis so that it is difficult to obtain enough mitotic chromosomes using a conventional colcemid block. Therefore, okadaic acid was used to condense prematurely the chromosomes of interphase cells. Even after irradiation with doses >40 Gy, okadaic acid was able to force sufficient numbers of cells to condense their chromosomes. Segments of chromosomes were then detected by chromosome painting using a goldconjugated antibody followed by silver enhancement. This simple method allows highly damaged chromosomes to be detected with great sensitivity.