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Electron Cryo-Microscopy

Key Facts

  

  • Single particle imaging for cryo-EM 3D reconstructions
  • Molecular and cellular electron cryo-tomography
  • Correlative light and electron cryo-microscopy

Multi-User Cryo-EM Facility

Electron Microscopes

The multi-user facility specialises in transmission electron microscopy of frozen biological samples.

Talos L120C (Thermo Fisher Scientific - FEI), 120 kV accelerating voltage, LaB6 source. This is a robust entry level screening microscope with a CETA camera, mainly for negative stain samples, but also capable of cryo-EM using side entry holders. SerialEM is installed on this microscope. This microscope is used for training purposes

Talos Arctica (Thermo Fisher Scientific FEI), 200 kV accelerating voltage, field emission gun (X-FEG) source. This microscope has an autoloader, Falcon 3EC direct detector and phase plates. This microscope is used primarily for cryo-EM sample optimization, single particle analysis and electron diffraction. A SerialEM installation is planned.

Titan Krios 1 (Thermo Fisher Scientific FEI), 300 kV accelerating voltage, field emission gun (X-FEG) source. Top-end microscope with autoloader, Bioquantum energy filter, K2 direct detector (to be upgraded to K3 in 12/2019), phase plate and dual axis tilt stage. This microscope is used for cryo-EM single particle and tomography data acquisition. SerialEM is installed.

Titan Krios 2 (Thermo Fisher Scientific FEI), 300 keV accelerating voltage, field emission gun (X-FEG) source. Top-end microscope with autoloader, Bioquantum energy filter, Falcon 3 EC (pre-filter) and K2 direct detector (to be upgraded to K3 in 09/2019), phase plate and single axis tilt stage. This microscope is used for cryo-EM single particle and tomography data acquisition. SerialEM is installed.

Aquilos (Thermo Fisher Scientific FEI) CryoDualBeam system dedicated to focused ion beam and scanning electron microscope (FIB-SEM) for the preparation of thin lamellae from vitreous cells for subsequent electron cryo-tomography. Uses autoloader compatible Autogrids.

 

Sample Preparation Equipment

In addition to the microscopes themselves, the electron microscopy equipment includes a broad range of auxiliary equipment and dedicated laboratory space for specimen preparation and mammalian cell culture.

  • Quorum GloQube glow discharger
  • Harrick Plasma Cleaner PDC-002
  • TFS Vitrobots Mark IV including one housed in a safety cabinet (Berner) for S2 sample preparation
  • Leica GP2 plunge freezer
  • MPI Martinsried type manual plunge freezers
  • Leica ACE60 Carbon/gold/platinum coater
  • Dedicated laboratory space for specimen preparation and mammalian cell culture
  • Safety Cabinets
  • CO2 incubators
  • Light Microscopes

Available Support

We provide a collaborative environment for sample preparation, data acquisition and data analysis. This includes:

  • structured user training
  • technical support for microscopes for state-of-the-art data acquisition
  • advice and collaborative development for sample preparation and project design
  • computing environment and guidance for initial data analysis
  • provision of various data processing pipelines
  • mutual development of user-specific workflows (both for data acquisition and processing)
  • regular user meetings and yearly cryo-EM workshops

Further information

We are a multi-user electron cryo-microscopy (cryo-EM) facility at the Center for Structural and Systems Biology (CSSB) on the Science Campus Bahrenfeld. The Multi-User CryoEM Facility at CSSB was established with substantial support from the DFG (grant numbers INST 152/772-1, 152/774-1, 152/775-1, 152/776-1 and 152/777-1 FUGG)

We are located at the Science Campus Bahrenfeld between Petra III and the EuXFEL. We aim to integrate and develop new methods between these facilities, e.g. micro-ED and electron crystallography. The facility operates under Biosafety S1 and S2 levels.

Facility access is managed by an online booking system (PPMS booking system coming shortly). Typically before starting a new project, an initial meeting will be arranged with the facility staff in order to discuss the plan and best suited approaches. Priority is given to projects that align with the scientific mission of the CSSB.

The cryo-CLEM technique is being arranged through our close interaction with the CSSB Advanced Light and Fluorescence Microscopy (ALFM) facility. We are in the process of developing and optimizing workflows for cryo-CLEM combining the fast detection of proteins inside cells by light microscopy with the high resolution of electron cryo-tomography. This is particularly powerful in mechanistically understanding host-pathogen interactions. Please contact us for more information on provisional access.

 

Cryo-EM Facility Team

4

Core Facility Head

Dr. Carolin Seuring
Phone:+49 040 8998 87707

E-Mail

Scientific Facility Director

Prof. Dr. Kay Grünewald

E-Mail

Staff Scientist (Computing)

Dr. Wolfgang Lugmayr
Phone:+49 40 8998 87652

E-Mail

Staff Scientist (Teaching)

Cornelia Cazey
Phone:+49 40 8998 87706

E-Mail

Location

CSSB Centre for Structural Systems Biology
c/o Deutsches Elektronen-Synchrotron DESY
Notkestraße 85, Building 15
D-22607 Hamburg

FACILITY OPERATOR

Universität Hamburg

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