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View All Electron Microscopy at Instruct
BREM offers services in high-resolution macromolecular cryo-EM and in-situ cryo-ET.
These services are enabled by Krios G4 300 kV and Talos F200i G2 200 kV cryo-TEMs, and Aquilos 2+ cryo-FIB-SEM. BREM has highly-qualified technical staff to run the facility and assist national and international users from academia and industry, under the guiding core principles of integrity, scientific excellence, collaboration, transparency, equal opportunity, diversity, and inclusion that align with those of INSTRUCT.
High-resolution macromolecular cryo-EM is enabled by a ThermoFisher Krios G4 300 kV cryo-TEM equipped with phase plate and paired with a BioContinuum energy filter and a K3 direct detector, and a Ceta-D CMOS camera for diffraction. This cryo-TEM is unique in southern Europe and is specially designed for all types of data collection at the highest possible resolution, including single-particle analysis (SPA), micro electron diffraction and cryo-ET. This service is also supported by a ThermoFisher iF200 G2 200 kV cryo-TEM, paired with a Falcon III direct detector and a Ceta-S CMOS camera, and a three-specimen Gatan Elsa cryotransfer system for specimen optimization. In addition, cryo-specimen preparation by immersion is enabled by two Vitrobot Mark IVs and a Leica EM Support preparation is enabled by a Q150VEplus carbon coater and high vacuum metallizer, an ELMO air glow discharger, and a Solaris II plasma cleaner.
In-situ cryo-ET of cells and tissues depends on the production of lamellas enabled by a ThermoFisher Aquilos 2+ focused cryo-FIB-SEM paired with an integrated epifluorescence microscope for cryo-CLEM. Imaging of lamellas by cryo-ET is performed in house on the Krios G4. In addition to cryo-specimen preparation by immersion, we can also carry out high-pressure freezing using a Leica EM ICE. In addition, we can offer tomographic data analysis services with our in-house software Dynamo (dynamo- em.org) developed by Daniel Castaño, which covers all steps from alignment of tilt series, reconstruction of tomograms, content characterization, identification of particles, subtomogram averaging and classification.
