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User Guide

CEMCOF is a cryo-EM user facility that is a part of CEITEC, Masaryk University. The facility is located at the University Campus Bohunice in Brno, Czech Republic and is organized by the specialists who support the facility users not only in cryo-EM data collection, but also in sample preparation, screening, and data analysis. The facility continuously adopts and further develops novel approaches and workflows in cryo-EM such as cellular lamella preparation by cryo-FIB or electron diffraction tomography (micro-ED) in order to increase the portfolio of the services provided to its users.

Provided services:

Our mission is to provide access to the state-of-the-art instrumentation for electron cryo-microscopy and support our user in sample preparation, data acquition, and data processing. Data analysis or training in different electron microscopy techniques are available through collaboration with CEMCOF. The services provided by the facility comprise:

• high-throughput single particle analysis (SPA) data collection
• cryo-electron tomography data acquisition
• cryo-FIB lamella preparation
• electron diffraction tomography (micro-ED) data acquisition
• data analysis
• sample preparation and cryo-EM screening
• resin embedding
• FIB-SEM tomography

The services are primarily provided using the following key instruments:

• Titan Krios – 300 kV high-end transmission electron microscope aligned for fringe-free imaging (FFI) and further equipped with Volta phase plate (VPP), energy filter, and Gatan K3 direct electron camera (Bioquantum 1067). The microscope serves for high throughput and high-quality single particle analysis and cryo-electron tomography data collection.
• Glacios 2 – 200kV (scanning-)transmission electron microscope for life-science applications. The microscope is equipped with a Falcon 4i direct electron detector for TEM applications, a Ceta-D camera for electron diffraction tomography, and a set of bright-field, dark-field and high angle annular dark-field detectors (Panther STEM, ThermoScientific) for STEM imaging. Data acquisition can be automated using SerialEM, EPU Multigrid and Tomo5 (including Tomo-Live for real-time tomogram reconstruction).
• Helios Hydra V – plasma-FIB/SEM dual beam microscope for room temperature and cryo-electron microscopy applications. High-resolution scanning electron microscope (Elstar column) with capacity for imaging in immersion mode and attainable resolution of 0.6nm
• Talso Actica – 200 kV high-end transmission electron microscope equipped with Ceta camera, Falcon 3EC direct electron detector, and post-GIF K2 direct electron detector (Bioquantum 967). The microscope serves for the acquisition of single particle analysis data, high-throughput screening of cryo-EM samples, and acquisition of electron diffraction tomography (microED) data. 
• Talos F200C – 200kV scanning transmission electron microscope for operation in both TEM and STEM modes, equipped with Falcon 3EC direct electron detector, Ceta-D camera, and Quadro (Dectris) direct electron detection camera.Talos F200C is a multi-purpose microscope for life-science and chemistry applications.
• Versa3D – small dual beam (FIB/SEM) equipped with cryo-stage and cryo-FIB/SEM preparation system (Quorum Technologies). The microscope primarily serves for the preparation of cellular lamellae by cryo-FIB for cryo-electron tomography.