Radiobiological studies using synchrotron-produced ultrasoft X-rays

Ultrasoft X-rays have been extensively used to explore radiobiological mechanisms surrounding cell killing. These studies for the most part have been linked to a small number of X-ray energies. Recently, this field of study has been broadened by the availability of synchrotron-produced ultrasoft X-rays which can be produced at any desired energy. We have taken advantage of the University of Wisconsin Synchrotron to reexamine two fundamental radiobiological questions: Dose RBE vary with different ultrasoft X-ray energies? Dose the fraction of the nuclear volume exposed to equal total X-ray energy modify cell cytotoxicity? The first study focuses on the survival of Chinese hamster V79 and mouse C3H10T1/2 cells irradiated with synchrotron-produced 273 eV and 860 eV ultrasoft X-rays. These two energies, which are available by multilayer monochromatization of the synchrotron output spectrum, exhibit equal attenuation within living cells. Such an isoattenuating energy pair allows the direct examination of how biological effectiveness varies with the energy of the ultrasoft X-rays. In comparing survival results, we find similar biological effectiveness of these two energies for both the C3H10T1/2 and the V79 cells. These results are no consistent with previous findings of increasing RBE with decreasing ultrasoft X-ray energies. In addition, after correcting for mean nuclear dose based on measurements of cell thickness obtained with confocal microscopy, we find no significant differences in survival between the two ultrasoft X-ray energies and 250 kVp X-rays. These results suggest that RBE does not increase with decreasing energy of ultrasoft X-ray between 860 eV and 273 eV. In a second study we introduced an method which allows partial-volume irradiation of live cells using synchrotron-produced ultrasoft X-rays and micro-fabricated irradiation masks. The masks were made by X-ray lithography at the University of Wisconsin Synchrotron Radiation Center, and they consist of 1.85-mu m-wide stripes of gold 1.35 mu m apart plated onto thin silicon nitrate membranes. When placed adjacent to mylar on which live cells are plated, these masks allow cells to be irradiated in a striped pattern with dimensions much smaller than the cell nuclei. Using 1340 eV synchrotron-produced X-rays, we compare the survival of cells subjected to uniform irradiation and cells subjected to partial-volume irradiation. Our results show that, at equal mean dose to the nucleus (i.e. equal total energies deposited), survival is not statistically different for the two treatments over a wide range of doses. Thus, imparting equal energies to smaller intranuclear volumes does not appear to modulate cell killing.