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RUPRECHT-KARLS-UNIVERSITÄT HEIDELBERG

Electron Microscopy Facility

 

Transmission electron microscopy (TEM) of biological macromolecular assemblies together with single-particle image processing is a powerful method for structure determination at intermediate resolution.
Information on the size, shape, homogeneity, and plasticity of macromolecular complexes or single proteins (≥ 200 kDa) can be achieved of samples that
exist in multiple identical copies. From such defined and reproducible arrangements a 3D structure at 1-3 nm resolution can be calculated. This allows the mapping of individual subunits or the modeling of high-resolution structures leading to a basis for functional understanding. (For an overview of the experimental steps see Böttcher and Hipp, 2010, "Single-particle applications at intermediate resolution.")

At the BZH we provide technical and scientific support for the complete workflow; from sample preparation for electron microscopy using e.g. the GraFix method, labeling of protein using e.g. the DID-Dyn2 label to the preparation of the negatively stained specimen on grids and a first quality check with an EM10 (Zeiss) microscope from Zeiss .
In close cooperation with the Electron Microscopy Core Facility (EMCF) at the Bioquant, we have unlimited access to microtomes, a freeze-substitution machine, high-end transmission EMs and a scanning EM as well as expertise in tomography and cryo-EM. This enables the groups of the BZH to answer a  variety of question from cell- to structural biology.



 










Fig: (left) Overview of an electron micrograph and class averages showing the dimerization of a reconstituted complex in a side-to-side arrangement.
Fig: (right) Three-dimensional reconstruction of a nucleoporin with revealing the structural similarity to karyopherins.


Contact:

 Dirk Flemming
BZH

Büro: +49 6221 54-4172


BZH

Labor: +49 6221 54-4166


BZH

Fax: +49 6221 54-5341


BZH

Electron Microscopy Facility


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