Electron Microscopy Core Facility
Model of meiotic SPB-Microtubule interaction in S.pombe cells from a tomography 3D reconstruction using IMOD (Univ. Boulder CO) (picture by Charlotta Funaya, collaboration with Kayoko Tanaka, University of Leicester)
Immunogold labelling of Cep152, a centriolar protein in HeLa cells (picture by Uta Haselmann, collaboration with Ingrid Hoffmann, German Cancer Research Centre (see Cizmecioglu et al., 2010))
The facility provides training in sample preparation and use of advanced electron microscopes, and develops approaches for cell and developmental biology.
The EMCF gives EMBL scientists the chance to learn sample preparation for EM and provides training on advanced electron microscopes and instrumentation, such as electron tomography (ET) setup (F30 300 kV TEM). Techniques can be applied and adapted to achieve EM-resolution at the level of cell organisation. We are also developing correlative microscopy approaches for cellular and developmental biology.
Major projects and accomplishments
Our electron tomography equipment, which has been operational since 2008, includes a new microscope and computing set-up with programs for 3D reconstruction and cellular modelling. The microscope is a FEI F30 (300 kV microscope with a field emission gun and Eagle FEI 4K camera) and is used mostly for cellular tomography of plastic-embedded samples, but can also be used as a cryo-microscope. Specialised EM engineers manage the F30, with expertise in tomography data acquisition and processing. Training is provided for researchers in handling the electron tomography microscope and its applications for cellular structure modelling.
We have upgraded our correlative light and electron microscopy (CLEM) technology with a new Zeiss LM microscope (Zeiss-Observer Z1), situated next to a high-pressure freezing machine (HPM-010) and devoted to perform CLEM, using cryofixation of our samples. New projects are welcome using this technology, and can be carried out in combination with methods developed at EMBL, such as by the Briggs group.
External collaborations include the study of SPB duplication in meiotic fission yeast (Kayoko Tanaka, University of Leicester); spindle formation in vertebrate cells (Maria Koffa, Democritus University of Thrace); SPBs in fission yeast (Anne Paoletti, Institut Curie-UMR144 CNRS); budding yeast SPBs (Schiebel group, DKFZ–ZMBH); and centriole biogenesis (Hoffmann group, DKFZ). We also support investigations on in vivo dengue virus replication (Bartenschlager group, University of Heidelberg).
Services provided
- Up-to-date knowledge of EM methods for cell biology, immunocytochemistry, cryosectioning and cryofixation applied to various cell types or organisms.
- Maintaining the microscopes and equipment for sample preparation, microtomy and cryogenic methods.
- Supplying a range of reagents specific to EM methods and protocols.
- Electron tomography, image acquisition and data processing for plastic-embedded samples.
- CLEM approaches and sample cryofixation.
- Assisting users in choosing the right methods and protocols.
- Organising courses and lectures on EM methods in cell biology.
Technology partners
FEI Company (advanced electron microscopes, including the new tomography microscope), Leica Microsystems (supplied our portable EMPACT2 HP Freezer, as well as ultramicrotomes units for plastic samples or cryo-sectioning) and Zeiss (for a light microscopy setup devoted to correlative LM/EM microscopy).