Serial femtosecond crystallography.

Area of Science:

• Structural Biology
• Biophysics
• Crystallography

Background:

• X-ray Free Electron Lasers (XFELs) enable novel macromolecular structure determination techniques.
• Serial crystallography methods like serial femtosecond crystallography (SFX) overcome limitations of traditional crystallography.
• SFX offers high throughput, reduced radiation damage, and time-resolved capabilities.

Purpose of the Study:

• To provide a practical guide for structural biology groups new to serial crystallography.
• To assist researchers in planning and executing successful SFX experiments.
• To highlight the potential and applications of SFX.

Main Methods:

• Microcrystal growth and characterization techniques.
• Sample delivery methods for high-throughput screening.
• Data processing and analysis pipelines for SFX data.

Main Results:

• SFX enables structure determination of challenging systems, including membrane proteins.
• Mitigation of radiation damage allows for studies of sensitive biological samples.
• Unprecedented temporal resolution for time-resolved structural dynamics.

Conclusions:

• SFX is a powerful technique for advancing macromolecular structure determination.
• This primer equips researchers with the knowledge to implement SFX.
• SFX opens new avenues for investigating biological processes at high resolution.