Real-time observation of irradiated HeLa-cell modified by fluorescent ubiquitination-based cell-cycle indicator using synchrotron X-ray microbeam

Narita, Ayumi; Kaminaga, Kiichi; Yokoya, Akinari; Noguchi, Miho; Kobayashi, Katsumi*; Usami, Noriko*; Fujii, Kentaro

For the knowledge about irradiation effects of mammalian cells depending on the cell cycle, most of them had been analyzed by statistical approches. Our purpose is to establish the method for real-time observation of irradiated cells under a microscope. Fluorescent ubiquitination-based cell cycle indicator (FUCCI) human cancer (HeLa) cells (red indicates G1; green, S/G2) were exposed to a synchrotron X-ray microbeam. Cells in either G1 or S/G2 were selectively irradiated according to cell color observed in the same microscopic field in a culture dish. Time-lapse micrographs of the irradiated cells were acquired for 24 h after irradiation. The cell cycle was strongly arrested by irradiation at S/G2 and never progressed to G1. In contrast, cells irradiated at G1 progress to S/G2 with a similar time course as non-irradiated control cells. These results show single FUCCI cell exposure and live cell imaging are powerful methods for studying radiation effects on the cell cycle.