Electron Microscopy(EM) has become an extremely popular method for the ultrastructural study of macromolecules, cells and tissues. An aqueous biological sample is frozen rapidly and irradiated with a beam of electrons. A detector senses how the electrons are scattered and a computer reconstructs the 3D-shape of the molecule.
Why do you need this service?
See the comparison of Crystallography, NMR, and EM
Service Processes: (Take protein as an example)
 Sample preparation |
After protein purification, these samples will be treated with cryo-fixation. In this method, protein samples are placed on a specially treated EM grid consisting of tiny holes in a film supported by a metal frame. The grid is then plunged into liquid ethane to flash-freeze it, resulting in the protein samples being embedded in a thin layer of vitreous ice. Once the frozen-hydrated grid is prepared, it is placed in the electron microscope and kept at approximately -180 K throughout the experiment. |
 EM imaging and data processing |
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 Model building and refinement |
Using our high-performance computers and software packages, we are able to interpret EM maps and reconstruct them into three-dimensional structural models that satisfy principles of physics and stereochemistry. (Our scientists are also very experienced with challenging targets such as membrane proteins by manual intervention to improve the initial fit or even build de novo models.) After an initial model is built, refinement is performed to maximize the agreement between the model and experimentally observed data by adjusting atomic coordinates, B factors, and other parameters. |
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