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Contenuto archiviato il 2024-05-29

Cryo- Electron Microscopy of vitreous sections of biological cells and tissues


During my postdoctoral research, I would like to investigate the macromolecular organization of synapses and desmosomes, in brain and skin tissue respectively, by combining cryo-sectioning (CS) and cryo-electron tomography (CET). This novel combination of CS with CET is currently the only approach which should allow the three-dimensional (3D) visualization of the organization of untreated (i.e. unfixed, unstained) tissue at macromolecular resolution.

The 3D molecular architecture of the synaptic cleft will help to understand the synaptic transduction processes. Revealing the 3D macromolecular organisation of the desmosomes and their associated structures (e.g. intermediate filaments) will have a great impact on understanding the mechanism by which desmosomes differentiate, and help to understand desmosome-related diseases (e.g. Pemphigus). In order to accomplish these goals, I will expand my expertise in CS with training in CET and image processing to be able to identify and analyse macromolecules in a cellular context. Tomographic series of cryo-serial sections are going to be recorded in the electron microscope followed by computationally reconstructing several 3D tomograms. Subsequently, the tomograms, of these sections will be computationally assembled in to a large 3D image which will represent a significant portion of the cellular volume at app. 2 nm resolution.

In the resulting reconstructions, I hope to reveal the molecular architecture of the desmosomes and synapses and thus we should be able to fit hi gh-resolution structures of adhesion proteins obtained by e.g. X-ray crystallography (e.g. Cadherin proteins in the desmosomes; neural cell adhesion molecules in the synaptic cleft). Finally I look forward to have strong interactions in the interdiscriplinary environment of EMBL, where I will also be able to supervise students and have knowledge-transfer in mutual directions. This should allow me to pave the way for a future leading position in science.

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