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Cryo-electron microscopy (cryo-EM) allows high resolution data of protein molecules, other macromolecules, macromolecular complexes or cells to be collected. In 2018 a grant of 30.8 mio DKK (4.7 mio USD) was allocated EMBION from the Ministry of Higher Education and Science – funded from 2015 call of the Danish Roadmap for Research Infrastrucutures. EMBION is a new cryo-EM consortium between Danish Universities (AU host and KU co-host). The National Cryo-EM Facility at AU (iNANO) started up medio 2019 (using existing electron microscopes and facilities). Based on the grant a new high-end electron microscope was purchased: a Titan Krios 300 keV transmission electron microscope with a Bioquantum/K3 detector setup and a Micro-ED system. Installation of the new Titan Krios (Krios2) microscope will finish in Q4 2020 and users can get access to the microscope from Q1 2021. The official inauguration of EMBION is 12th October 2020.

Fully equipped EMlab

An EMlab with the equipment necessary for screening and optimizing samples for negative stain and cryo-EM single particle analysis and other biological EM work is part of the Facility. This includes a ThermoFisherScientific Vitrobot Mark IV, a Leica EM GP2 Automatic Plunge Freezer, Leica EM SCD 500 High Vacuum Sputter Coater, glow dischargers (EasiGlow and GloQube), Leica CPD 300 Automated Critical Point Dryer, Leica EM UC7 Ultramicrotome and Fischione Model 1020 Plasma Cleaner. Furthermore, based on a recent grant from the Novo Nordisk Foundation it will be possible in the near future to prepare samples for cryo-electron tomography using focused ion beam milling under cryogenic conditions - during 2021 a cryo-FIB/SEM and a high pressure freezer will be installed. In Q4 2020 the plungers will be moved to a humidity (RH of <35%) and temperature (22°C) controlled lab next to Krios2.

Automated data collection

High resolution single particle data can be collected automatically on Titan Krios 1 and 2 using EPU and high resolution tomography data can be collected automatically using SerialEM. The K2/K3 PCs are connected with dedicated optical cables to our local petabyte data storage and high performance computing cluster (Electron Microscopy Computing Cluster - EMCC) allowing ultrafast data transfer and processing. Data is streamed using the Scipion or Warp pipelines which give the user real time information on data quality and allows the first steps of data processing to be performed in parallel to data collection. Access to EMCC is available based on a yearly fee - please contact scientific computing specialist Jesper L. Karlsen (jelka@mbg.au.dk) for information. Data will be stored on EMCC for a certain period (currently 3 months) and will be handed out to the user on an external harddrive.

Use of the facility

The Facility offers local AU users, users from other DK research institutions, international academic users (through the iNEXT Discovery program) and industrial users access to state of the art cryo-EM equipment and specialist assistance in a range of areas including single particle analysis and tomography. In order to apply for access new users need to register. Users can apply for introductory training in TEM by contacting EM Tecnical Manager Ramon Liebrechts (rlie@inano.au.dk) or obtain information on access to the high-end microscopes by contacting Cryo-EM Facility Manager Thomas Boesen (thb@mbg.au.dk). Alternatively, use the contact form on this web portal for inquiries.

Workhorse Tecnai 120 keV microscopes

It is important to screen and optimize samples in order to document that the project is at a stage ready for getting access to the high-end Titak Krios microscopes. This screening and optimization can be done on the 120 keV Tecnai Spirit transmission microscope. Single particle data from negative stain samples can be collected automatically with Leginon. The data are stored in a database for on the fly processing using Appion or another pipeline. In addition SerialEM is installed on the Tecnai Spirit. In Q4 of 2020 a second 120 keV Tecnai microscope dedicated cryo-EM screening will be installed at the facility.

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