200 keV FEG Electron Cryo Microscope

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2017
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 342247113

Highest resolution structural biology analyses of macromolecules require optimal sample preparation. Besides a full biochemical characterisation this optimisation includes particularly the cryo preparation (vitrification) itself that requires a systematic iterative approach guided by feedback from preliminary EM analyses. In this optimisation e.g. various blotting conditions are tested to ensure best preservation, ultimate sample orientation and ice condition. This screening includes acquisition of data sets for preliminary processing, to allow better sample characterisation with respect to conformational states. A 200keV FEG electron cryo microscope with direct electron detector and robotic Sample changer fulfils these requirements. Given the already foreseeable oversubscription of the Titan Krios for single particle cryoEM, this microscope will ensure a good workflow and that only well-characterised samples and matured projects move on to the top-end instrument. Further, the instrument will serve the needs of molecular cryoET of thinner specimens and initial characterisation of the specimens coming off the cryoFlB-SEM. The possibility to use the common autogrid sample format with the high-end instruments allows efficient sample progression across the microscopes and enables users to gain experience with handling all the major functionality of the high-end instruments. Systematic sample screening and transfer success is significantly higher on autoloader than in ‘side-entry' systems. Putting the EM in a S3 area, further allows for analyses of infectious material. Together with the top-end instruments a very wide range of samples is covered.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: 200 keV FEG electron cryo microscope
• Instrumentation Group: 5100 Elektronenmikroskope (Transmission)
• Applicant Institution: Universität Hamburg
• Leader: Professor Dr. Kay Grünewald