X-ray Free Electron Lasers (XFELs) have advanced research in structure biology, by exploiting their ultra-short and bright X-ray pulses. The resulting “diffraction before destruction” experimental approach allows data collection to outrun radiation damage, a crucial factor that has often limited resolution in the structure determination of biological molecules. 

Since the first hard X-ray laser (the Linac Coherent Light Source (LCLS) at SLAC) commenced operation in 2009, serial femtosecond crystallography (SFX) has rapidly matured into a method for the structural analysis of nano- and micro-crystals. At the same time, single particle structure determination by coherent diffractive imaging, with one particle (such as a virus) per shot, has been under intense development. In this review we describe these applications of X-ray lasers in structural biology, with a focus particularly on aspects of data analysis for the computational research community.We summarize the key problems in data analysis and model reconstruction, and provide perspectives on future research using computational methods.

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