From solution to structure: empowering inclusive cryo-EM with a pre-characterization pipeline for biological samples

Area of Science:

• Structural Biology
• Biophysics
• Microscopy

Background:

• Limited access to cryo-electron microscopy (cryo-EM) infrastructure hinders research in many academic institutions.
• There is a need for accessible, high-throughput cryo-EM services for structural determination.

Purpose of the Study:

• To introduce the cryo-EM Solution-to-Structure (SOS) pipeline.
• To provide a comprehensive service for researchers lacking in-house cryo-EM facilities.
• To enable high-resolution structural analysis through single-particle analysis and cryo-electron tomography.

Main Methods:

• The SOS pipeline includes grid preparation, negative stain imaging (for protein samples), grid screening, and preliminary data collection.
• It offers two entry points: submission of purified protein samples or already vitrified grids.
• High-resolution data collection is performed on a Titan Krios microscope with a direct electron detector and energy filter.

Main Results:

• The pipeline successfully integrates sample preparation, screening, and high-resolution data collection.
• It accommodates both single-particle analysis and cryo-electron tomography workflows.
• Successful completion of screening steps leads to high-resolution data acquisition.

Conclusions:

• The SOS pipeline effectively democratizes access to cutting-edge cryo-EM technology.
• It offers a robust solution for researchers to achieve high-resolution structures.
• The pipeline streamlines the process from sample to structure, enhancing research capabilities.