Carbon evaporation system for preparation of electron microscopy grids

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Cryo-electron microscopy (cryo-EM) has contributed majorly to our knowledge of macromolecules performing critical functions in the cell. Understanding the structure of these protein complexes is essential to overcome major threats to human health such as bacterial antibiotic resistance or cancer. In cryo-EM, the specimen is embedded in an amorphous form of ice called "vitreous ice" in which the specimen remains fully hydrated. Vitrification is performed in buffer conditions that are physiologically relevant for the specimen. Therefore, cryo-EM pictures constitute a snapshot of a physiologically relevant state of the macromolecular assembly from which a 3D model is calculated. The molecular mechanisms of macromolecules are then inferred form the obtained three-dimensional structures. Prior to the vitrification process the sample is usually deposited in an electron microscopy grid that has been previously coated with a thin layer of carbon. A high-quality carbon coated grid is essential for the successful vitrification of the sample that precedes the high-resolution imaging of the macromolecules in an electron microscope. The requested equipment is a carbon evaporation system designed to reliably deposit high quality carbon films in electron microscopy grids. The requested equipment will streamline our sample preparation procedures and will allow us to efficiently prepare samples for cryo-EM. This equipment will also be used for cryo-EM applications in materials research performed by Dr. Gianluigi Botton, a co-applicat in thi proposal, and other Canadian and foreign visiting scientist of the Canadian Centre for Electron Microscopy at McMaster University. Advancing our research program in structural biology will certainly increase our chances to develop strategies to fight diseases that are a major threat to human health. In addition, the use of this instrument for material research will contribute to the development of new polymers and carbon nanostructures. Finally, state-of-the-art instrumentation translates onto first-class training. The requested equipment will help to attract and retain the best trainees in Canada and also to provide them with the best possible training experience.