National Facility for Structural Biology

National Facility for Structural Biology

The National Facility for Structural Biology (StructBio) provides a comprehensive platform for structural characterization across scales, from tissue to amino acid side-chains. The NF StructBio will be operated by highly trained technologists with expertise in all aspects of sample preparation, specimen characterization and imaging. The Facility aims at supporting the national scientific community in successful investigating the biological players of interest, both isolated, and in their cellular compartment.

The NF for StructBio foresees six Infrastructural Units (IU):

• INFRASTRUCTURAL UNIT 1 Cryo-Electron Microscopy
• INFRASTRUCTURAL UNIT 2 Biomass Production
• INFRASTRUCTURAL UNIT 3 Biophysics
• INFRASTRUCTURAL UNIT 4 Structural Proteomics
• INFRASTRUCTURAL UNIT 5 Dynamic Single-molecule
• INFRASTRUCTURAL UNIT 6 Technology Development

Available equipment

To ensure maximum productivity and research quality, the NF for StrucBio benefits from state-of-the-art equipment. The list of technologies available for external user to access the Cryo-Electron Microscopy Unit (IU1) includes:

• a Thermo Scientific Titan Krios G4i 300kV TEM equipped with Thermo Scientific Falcon 4i 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 4i, Thermo Scientific Selectris X energy filter, and CETA-D 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 Leica Stellaris 5 confocal microscope equipped with white-light laser and cryo-stage to perform cryo-CLEM experiments;
• a Leica Thunder widefield microscope equipped with cryo-stage to perform cryo-CLEM experiments;
• ancillary equipment for sample preparation including: plunge freezing devices (Thermo Scientific Vitrobot Mark IV), glow discharger units (Pealco EasyGlow and Quorum GloQube), plasma cleaner (Gatan Solarus II), carbon coating system (Leica ACE600), and other sample preparation tools for high-pressure freezing (Leica EM ICE), and freeze substitution (Leica AFS2).

Biomass Production & Biophysics Unit (IU2) will offer, among other dedicated instruments:

• a fermentation room equipped withfermenters for large scale production in yeast and bacteria (InforsHT Techfors-s 15L and Techfors 150L);
• insects and mammalians cell production room with two bioreactors of 0,5L (myControl) and 15L (ezControl) ;
• instruments for sample characterisation:
  • Refeyn OneMP (Mass Photometry);
  • Xtal Concepts SpectroLight 610 (Dynamic Light Scattering);
  • Nanotemper Tycho NT.6 and Prometheus NT.48 (nanoDSF)
• instruments for measurement of affinities:
  • Microcal PEAQ-ITC (Isothermal Titration Calorimetry;
  • Sartorius Octet R8 (BioLayer Interferometry);
  • Nanotemper Monolith (Microscale Thermophoresis)