Carbon evaporation system for preparation of electron microscopy grids

冷冻电子显微镜（cryo-EM）的主要贡献，我们的知识大分子在细胞中执行的关键功能。了解这些蛋白质复合物的结构对于克服对人类健康的主要威胁（如细菌抗生素耐药性或癌症）至关重要。在冷冻EM中，样本被嵌入称为“玻璃冰”的无定形冰中，其中样本保持完全水合。在与标本生理相关的缓冲液条件下进行玻璃化。因此，冷冻EM图片构成了大分子组装体生理相关状态的快照，从中计算出3D模型。然后从所获得的三维结构推断大分子的分子机理。在玻璃化过程之前，通常将样品沉积在预先涂覆有薄碳层的电子显微镜网格中。高质量的碳涂层网格对于在电子显微镜中高分辨率成像之前成功玻璃化样品至关重要。所要求的设备是一个碳蒸发系统，旨在可靠地存款高质量的碳膜在电子显微镜网格。所要求的设备将简化我们的样品制备程序，并使我们能够有效地制备冷冻EM样品。该设备还将用于材料研究中的低温EM应用，由本提案的共同申请人Gianluigi Botton博士以及麦克马斯特大学加拿大电子显微镜中心的其他加拿大和外国访问科学家进行。推进我们的结构生物学研究计划肯定会增加我们制定战略的机会，以对抗对人类健康构成重大威胁的疾病。此外，将该仪器用于材料研究将有助于开发新的聚合物和碳纳米结构。最后，最先进的仪器转化为一流的培训。所要求的设备将有助于吸引和留住加拿大最好的受训人员，并为他们提供尽可能好的培训经验。

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.