Serial femtosecond crystallography: the first five years.

Area of Science:

• Structural Biology
• Biophysics
• Biochemistry

Background:

• Protein crystallography using synchrotron radiation has advanced biological understanding but is limited by crystal size and radiation damage.
• Cryogenic temperatures are typically used to mitigate radiation damage during data collection.

Purpose of the Study:

• To review novel approaches for crystal growth, delivery, and diffraction data analysis in serial femtosecond crystallography (SFX).
• To describe the opportunities and challenges of SFX, including its application in time-resolved studies and radiation-sensitive systems.

Main Methods:

• Serial femtosecond crystallography (SFX) utilizes intense X-ray pulses from free-electron lasers (FELs).
• Microscopic crystals are exposed to femtosecond X-ray pulses, yielding diffraction data before destruction.
• Data are collected in a serial manner from numerous crystals at cryogenic or room temperature.

Main Results:

• SFX allows for the structure determination of undamaged macromolecules, even from microscopic crystals.
• New methods for crystal handling, delivery, and de novo phasing are essential for SFX data analysis.
• The technique facilitates time-resolved experiments and the study of radiation-damage-prone biological samples.

Conclusions:

• SFX offers a powerful alternative to traditional crystallography, overcoming crystal size and radiation damage limitations.
• This method opens new avenues for structural biology, particularly for challenging biological systems.
• Further development in experimental and analytical approaches will expand the utility of SFX.