Electron Microscopy Core Facility
EM view of a synapse at the neuromuscular junction in mouse skeletal muscle (Uta Haselmann, EMBL with Anna Gail, DKFZ).
The EMCF gives EMBL scientists access to advanced electron microscopes, relevant sample preparation techniques and specialised instrumentation, in particular the newly installed electron tomography set-up. Techniques can be applied and adapted to various projects across the units to access EM resolution at the level of cell organisation. The facility also trains new users to make best use of our advanced equipment and develops new approaches and methods in EM applications to cellular and developmental biology.
Major projects and accomplishments
Our new Electron Tomography equipment includes a new microscope and computing set-up with effective programs for 3D reconstruction and image modelling. Mostly destined to perform cellular tomography of plastic embedded samples, the new microscope is a FEI F30 (300 kV microscope with a Field Emission Gun and Eagle FEI 4K camera). It is also equipped as a cryomicroscope to support cryo-EM investigations. This microscope is managed by specialised EM engineers who hold the knowledge for tomography data acquisition and processing, teach users the various applications for cellular structure modelling and introduce them to the F30 microscope.
Correlative microscopy technology (a collaboration between EMCF and ALMF, page 55) has been established with conventionally fixed cells grown on coverslips (Colombelli et al., 2008, Methods in Molecular Biology, in press). We are now developing a similar method adapted to cells grown on sapphire coverslips which are destined to be cryofixed by high-pressure freezing after LM visualisation.
Other projects include investigations on annexins in plasma membrane-injured cells (Schultz group, page 36); a p53 network built in budding yeast (Barbara di Ventura, Serrano Group, formerly EMBL Heidelberg; Di Ventura et al., 2008); and morphology of synapse in wt and mouse tumour (Anna Gail, Molecular Metabolic Control, Stephan Herzig group, DKFZ, Heidelberg).
Services provided
- An up-to-date know-how on EM methods for cell biology, immunocytochemistry, cryosectioning and cryofixation applied to various cell types or organisms.
- Maintaining the electron microscopes and the equipment in the laboratory for sample preparation, microtomy and cryogenic methods.
- Supplying a range of reagents specific for the relevant EM methods and protocols.
- Electron Tomography, image acquisition and data processing for plastic embedded samples.
- Assisting users in choosing the right methods and protocols for their project.
- Organising courses and lectures on EM methods in cell biology.
Technology partners
- FEI Company: Supplier of advanced electron microscopes, including the new tomography microscope.
- Leica-microsystems is the constructor of our HPFreezer EMPACT2, a portable ma chine with an optional attachment, the Rapid Transfer System (RTS), which per mits easy loading of the samples and allows correlative light and electron microscopy. They are also our supplier for the various ultramicrotomes units we use for sample plastic- or cryo-sectioning.