Single Particle Cryo-Electron Microscopy: From Sample to Structure

Cryo-electron microscopy (cryoEM) is a powerful technique for structure determination of macromolecular complexes, via single particle analysis (SPA). The overall process involves i) vitrifying the specimen in thin film supported on cryoEM grid; ii) screening to assess distribution and ice quality; iii) if grid suitable, collecting dataset analysis; iv) image processing yield an EM density map. In this protocol, overview each these steps provided, with focus variables which user can modify during workflow troubleshooting common issues. With remote microscope operation becoming standard many facilities, variations imaging protocols assist users efficient when physical access limited will be described.