Effect of crystal-to-detector distance shift on data processing in serial crystallography.

Area of Science:

• Structural Biology
• Biophysics
• Crystallography

Background:

• Serial crystallography (SX) enables room-temperature macromolecular structure determination with minimized radiation damage.
• Continuous crystal delivery in SX leads to variations in crystal-to-detector distance (CTDD) during data collection.
• The impact of CTDD variations on SX data quality and processing remains incompletely understood.

Purpose of the Study:

• To assess the influence of crystal-to-detector distance (CTDD) accuracy on serial crystallography (SX) data quality.
• To investigate how variations in CTDD affect data processing parameters and structural results.
• To provide insights for optimizing SX data processing and interpretation.

Main Methods:

• Collected serial synchrotron crystallographic data for hen egg white lysozyme (HEWL) and glucose isomerase (GI).
• Processed SX data using three indexing algorithms at various CTDDs.
• Analyzed unit cell distributions, indexing trends, and structure refinement statistics (Rfree) at different CTDDs.

Main Results:

• The CTDD yielding maximum indexed images differed from the optimized CTDD, with algorithm-dependent trends.
• Deviations from the actual CTDD distorted unit cell distribution and altered indexed unit cell dimensions.
• Structure solution was achievable with ±10 mm CTDD deviation for HEWL and GI, though data quality decreased with larger deviations.

Conclusions:

• CTDD accuracy significantly impacts SX data processing and unit cell parameter determination.
• While some CTDD deviation is tolerated for structure solution, optimal data quality requires precise CTDD.
• These findings are valuable for improving the efficiency and reliability of serial crystallography data processing.