Electron Microscopy Core Facility
Meiotic SPB-Microtubule interaction in S.pombe cells from a serial section 3D reconstruction and modelling using IMOD (Univ. Boulder CO). Picture by C. Funaya, collaboration with K. Tanaka.
Immunogold labelling of Cep152, a centriolar protein in Hela cells. Picture by U. Haselmann, collaboration with I. Hoffmann J Cell Biol. 191:731-9.
The EMCF gives EMBL scientists the opportunity to learn sample preparation for EM and provides training on advanced electron microscopes and specialised instrumentation, in particular the electron tomography setup. These techniques can be applied and adapted to various projects across the different units to achieve EM resolution at the level of cell organisation. The facility is also developing correlative microscopy approaches for cellular and developmental biology questions..
Major projects and accomplishments
Our electron tomography equipment, 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. Nevertheless, it can also be used as a cryo-microscope. The F30 is managed by specialised EM engineers 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. In 2010 we welcomed Rachel Mellwig, EMCF operations manger, as a new staff member.
Correlative Microscopy technology has been a fruitful approach to study the biogenesis of the Golgi apparatus (see Tangemo et al 2011). 3D reconstruction of the dorsal closure in the Drosophila embryo (Brunner and Frangakis groups, formerly EMBL) and the microtubule based polarity of the Drosophila oocyte (Ephrussi group) are ongoing projects based on EM investigations involving electron tomography. External collaborations include the study of SPB duplication in meiotic fission yeast (K. Tanaka, University of Leicester, UK), bacterial protein expression in fission yeast (M. Balasubramanian, Singapore), and in vivo dengue virus replication (R. Bartenschlager group, University of Heidelberg). We have also looked at SPBs and centrioles using immunolabelling techniques: SPB-microtubule interactions in budding yeast (E. Schiebel group, DKFZ-ZMBH, Heidelberg) and centriole duplication (I. Hoffmann group, DKFZ, 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 equipment for sample preparation, microtomy and cryogenic methods.
- Supplying a range of reagents specific for EM methods and protocols.
- Electron tomography, image acquisition and data processing for plastic-embedded samples.
- Assisting users in choosing the right methods and protocols.
- Organising courses and lectures on EM methods in cell biology.
Technology partners
- FEI Company
- Leica Microsystems
- Zeiss
- Martin Wohlwend GmbH (CH)