Detection of DNA Damage in Individual Cells Induced by Heavy-ion Irradiation with an Non-denaturing Comet Assay

Investigating the biological effects of high-LET heavy-ion irradiation at low fluence is important to evaluate the risk of radiation in space. It is especially necessary to detect radiation damage induced by a precise number of heavy ions in individual cells. We thus compared the number of ions traversing a cell and the DNA damage produced by ion hits. We applied a comet assay to measure the DNA damage in individual cells. Cells attached on ion-track detector (CR-39) were irradiated with 17.3 MeV/u 12C, 15.7 MeV/u, 10.4 MeV/u 20Ne ion and 7.2 MeV/u 40Ar beams at TIARA, JAERI-Takasaki. After irradiation, CR-39 was covered with 1% agarose. The agarose was allowed to solidify on a glass slide, and then the electrophoresis was performed. Afterward, the CR-39 was taken off the glass slide. The agarose gel on the CR-39 was stained with ethidium bromide and the opposite side of the CR-39 was etched with a KOH-ethanol solution at 37 degrees C. We observed that heavy ions with higher LET values induced heavier DNA damage, even with the same number of ion hits within the irradiated cells. The result indicated that the amount of DNA damage induced by one particle depended on the LET value of the heavy ions.