Experimental Method For Researching The Radiation Damage Effects On Single Cell By Soft X-ray

When the living cell was irradiated by X-ray with some doses, the DNA in cellswill be damaged and even result in the cell mutation or death. Studying the radiationdamages and the biological effects in irradiated cells, will provide a theoretical basisfor radiation therapy, risk assessment and radiation protection. In conventionalresearch, due to the limitation of methods, groups of cells were irradiated and tested,instead of a single cell. Therefore, the result of injury effect is an average effect ofgroup rather than single cell. It is necessary to develop the method for researching theradiation damages in single cell.In this study, the experimental device was set up based on the soft X-raymicroscopy beamline station in Hefei Light Source (HLS). The soft X-ray wasfocused into micro-beam by a linear monochromator (constitutes of a condenser zoneplate and a pinhole), and the energy calibration and spot determination were carriedout. The irradiate time was controlled to ensure the irradiated cell receive an accurateradiation dose, according to the current beam intensity of synchrotron radiationstorage ring.The cell culture system and method for the single cell irradiation was formed toculture the Hela cells which are selected as the sample in the experiments. Thelocating and environment maintaining system were designed to ensure the normalphysiological cell activity.Single Cell Gel Electrophoresis (SCGE) and Synchrotron Radiation FourierTransform Infrared (FTIR) microspectroscopy were used for detecting the irradiatedcells. The results suggest that SCGE can detect the DNA strand breaks sensitively, andcan show that the radiation damages has a dose-dependent tendency. SynchrotronRadiation FTIR microspectroscopy can accurately detect the radiation effects in cells,such as DNA strand breaks, purine / pyrimidine methylation, and so on.In summary, our work provides a novel useful exploration for researching thebiological effects in single irradiated cell, and expands the new application of FTIRmicrospectroscopy for detecting the irradiation damage in single cell.