The facility will provide support and expertise to researchers with samples in the first stages of characterization. Samples will be analysed by negative staining in search of the best conditions. Once found, samples will be subjected to different vitrification conditions in search of the best parameters, which will be tested in a cryomicroscope Talos Arctica 200 kV equipped with a Falcon III electron direct detector.
The best grids will be used to acquire data for image processing (it is advisable to contact the Instruct Image Processing Centre for support in the data processing).
Based at the premises of the Centro Nacional de Biotecnología (CNB), the CSIC cryoelectron microscopy (CryoEM) facility is a state-of-the-art core service that offers both sample preparation and CryoEM image collection of unstained biological material.
The facility hosts two cryoelectron microscopes: A 300 kV JEOL CryoARM equipped with an autoloader, a Gatan K3 direct electron detector and an Omega energy filter, and a 200 kV FEI TALOS Arctica, equipped with an autoloader and a Falcon III direct electron detector. Both are suited for the collection of large amounts of high-resolution data that can be used for high-resolution studies using single-particle methodology, or for the reconstruction of pleiotropic structures such as cells and subcellular structures using cryoelectron tomography (cryoET). Additionally, the facility hosts a standard 120 kV electron microscope JEOL JEM1400 for sample screening.
Specimen vitrification is also available at the service, with two devices suited for vitrification of small specimens such as proteins and macromolecular complexes (a FEI Vitrobot and a Leica EM CPC), and a third device suited for the fast-freezing of cellular samples, a Leica EM GP2 high-pressure freezer. This facility is unique in Spain as it also offers the newly developed cryocorrelative microscopy technique, which allows the analysis of the same biological specimen by cryooptical microscopy (Zeiss LS900 AiryScan cryoconfocal microscope) and cryoelectron microscopy (both cryoelectron microscopes described above). The use of a Zeiss CrossBeam 550 cryo-FIB-SEM microscope will increase the cryocorrelative microscopy capabilities of the facility to direct visualization of cells for tissue-cell resolution or for preparation of thin lamellas in cells for molecular resolution.
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