Cryo-Electron Microscopy FacilityHEAD: Paolo Swuec
Why a Cryo-Electron Microscopy Facility?
The Cryo-Electron Microscopy Facility’s mission is to provide access to a highly productive, world-class scientific hub, capable of solving at high resolution, from tissue to amino acid side-chains, the many questions faced by contemporary life science.
Every biological process, including physiologic and pathologic events, is precisely orchestrated by active and reactive biological macromolecules. The function, organisation and activity of these molecules strictly depend on both their three-dimensional (3D) structure and the cellular environment in which they operate.
The Cryo-EM Facility at Human Technopole aims at identifying, visualising, and characterising these biological players alone and in their cellular compartment.
Key features of the Cryo-Electron Microscopy Facility
To ensure maximum productivity and research quality, the Cryo-Electron Microscopy Facility benefits from state-of-the-art equipment including:
- A Thermo Scientific Titan Krios G4i 300kV TEM equipped with Thermo Scientific Falcon 4 direct electron detector, Thermo Scientific Selectris X energy filter, Thermo Scientific CETA 16M and Volta phase-plate
- a Thermo Scientific Spectra 300kV STEM equipped with a Thermo Scientific CETA 16M with speed enhancement fully dedicated to electron tomography workflows
- a Thermo Scientific Glacios 200kV TEM equipped with Thermo Scientific Falcon 4 and CETA 16M and Volta phase-plate
- a Thermo Scientific Talos L120C 120kV TEM equipped with Thermo Scientific CETA 16M to allow both room temperature and cryogenic imaging (by Gatan ELSA cryo-holder)
- a Thermo Scientific Aquilos 2 DualBeam cryo-FIB system.
Ancillary equipment includes a fluorescence microscope with a cryo-stage for CLEM applications, plunge freezing devices, glow discharger units, plasma cleaners, carbon coating system, and other sample preparation tools for high-pressure freezing, freeze substitution and ultramicrotomy of resin embedded and vitrified samples.
Examples of experiments that can be carried out at HT Cryo-EM Facility
Human Technopole’s Cryo-Electron Microscopy Facility is designed to efficiently combine single-particle analysis (SPA), electron tomography (ET) and correlative light electron microscopy (CLEM) workflows to study in detail the structure of single macromolecules as well as whole cellular compartments. The Cryo-EM Facility will support scientific demand of HT’s five interdisciplinary research centres. For example, by SPA it is possible to obtain the high-resolution 3D structure of proteins, enzymes and other macromolecules. Also, ET is employed at the Cryo-EM Facility to explore the inner architecture of cells, tissues and organoids. By means of focused ion beam – scanning electron microscopy (FIB-SEM) technology the Cryo-EM Facility can open a window to look inside vitrified samples. Beside all of these, we support and carry preliminary sample preparation (e.g. high-pressure freezing, freeze-substitution, heavy metal staining, plunge freezing, micro-patterning) and investigations (e.g. sample screening by negative staining and Cryo-EM, correlative microscopy).