Single-cell Gel Technique Supports Hit Probability Calculations

Abstract

We have used the alkaline single-cell gel technique to provide a biological estimate of the percentage of cell nuclei "hit" by alpha particles during in vitro radon exposure. The single-cell gel electrophoretic technique measures DNA strand breaks as increased migration of the DNA out of lysed cells embedded in the middle layer of a three-layer gel formed on a microscope slide. Two of the advantages of this system are that individual cells of an exposed population can be evaluated and that histograms can be constructed to estimate the population response. Chinese hamster ovary and AL cells were each exposed to 0.39 Gy of radon, a dose at which our dosimetry model predicts that 63 and 73%, respectively, of the cell nuclei will be traversed by an alpha particle. The difference in the percentages at similar doses is mainly due to the larger nucleus volume in AL cells. A 1.5-Gy x-ray response was also evaluated as a low-LET control. As expected, the x-ray profile of DNA damage was shifted from the nonirradiated profile in the direction of greater DNA migration and approximated a normal distribution. The profile of the radon-exposed cells was biphasic, with one distribution corresponding to the control (nonirradiated) response and the other profile showing increased DNA migration. We interpret the second profile in the biphasic profile as representing cell nuclei that had received an alpha "hit." The percentages of cell nuclei in the "hit" category (approximately 51 and 45% for CHO and AL, respectively), as judged by the single-cell gel technique, were 81 and 62% of the calculated values.