Some practical considerations about the effects of radiation damage on hydrated cells imaged by X-ray fluorescence microscopy

X-ray fluorescence (XRF) microscopy features unique capabilities which make it well suited for biological investigations. Its high sensitivity together with high spatial resolution and penetration depth provide a unique tool for trace elements analysis in heterogeneous samples. Like most of the X-ray based techniques, radiation damage sets hard limits on the ultimate performance. Although the interactions between matter and photons are well described from a physics point-of-view, there is a lack of experimental data, in particular for XRF imaging mode. In this context, this work proposes a practical approach in addressing the limits set by radiation damage to X-ray fluorescence imaging in the case of hydrated and unfixed cells at room temperature. We find that the maximum dose tolerated by ascidian blood cells is 10⁵ Gy. A simple theoretical model allowed the minimal doses required for a good image contrast to be determined for various experimental schemes. The results are consistent with the experimental observation on ascidian blood cells which exemplifies the peculiar case of highly concentrated samples (>10,000 ppm) at room temperature. The same simple model predicts that in the case of the detection of high Z trace elements in cryo-preserved cells, the relative detection limit set by radiation damage is below 0.1 ppm at a spatial resolution of 100 nm.