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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.
The Workflow of Our Cryo-EM Services (Take protein as an example)
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|
|Model building and refinement|
Using our high-performance computers and software packages, we can interpret EM maps and reconstruct them into 3D structural models that satisfy the 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.
Creative Biostructure has extensive experience in using cryo-EM technology to study dynamic conformational changes and function of macromolecular complexes and has successfully applied this technology to characterize the structures of gene therapy and drug delivery particles. If you are interested in our cryo-EM services, please feel free to contact us to discuss your project.