Serial femtosecond crystallography

With the advent of X-ray free-electron lasers (XFELs), new, high-throughput serial crystallography techniques for macromolecular structure determination have emerged. Serial femtosecond crystallography (SFX) and related methods provide possibilities beyond canonical, single-crystal rotation crystallography by mitigating radiation damage and allowing time-resolved studies with unprecedented temporal resolution. This Primer aims to assist structural biology groups with little or no experience in serial crystallography in planning and carrying out a successful SFX experiment. It discusses the background of serial crystallography and its possibilities. Microcrystal growth and characterization methods are discussed, alongside techniques for sample delivery and data processing. Moreover, it gives practical tips for preparing an experiment, what to consider and do during a beam time and how to conduct the final data analysis. Finally, the Primer looks at various applications of SFX, including structure determination of membrane proteins, investigation of radiation damage-prone systems and time-resolved studies.

Automated summary

This article introduces the basic principles of X-ray free-electron lasers and serial crystallography, providing assistance for structural biology groups in planning and conducting SFX experiments. The main content includes microcrystal growth and characterization, sample delivery and data processing techniques, experimental preparation and operation at different stages, as well as the possibilities of SFX in various application areas.