Carbon evaporation system for preparation of electron microscopy grids

低温电子显微镜(Cryo-EM)为我们了解大分子在细胞中发挥关键功能做出了主要贡献。了解这些蛋白质复合体的结构对于克服细菌抗生素耐药性或癌症等对人类健康的主要威胁至关重要。在低温EM中，标本被嵌在一种被称为“玻璃冰”的无定形冰中，在这种冰中，标本保持充分的水合。玻璃化冷冻是在与标本生理相关的缓冲条件下进行的。因此，冷冻-EM图像构成了计算3D模型所依据的大分子组件的生理相关状态的快照。然后从所获得的三维结构中推断出大分子的分子机制。在玻璃化过程之前，样品通常沉积在电子显微镜网格中，该网格之前已经覆盖了一层薄薄的碳层。在电子显微镜中对大分子进行高分辨率成像之前，高质量的碳涂层网格对于样品的成功玻璃化至关重要。所要求的设备是碳蒸发系统，旨在可靠地在电子显微镜栅格中沉积高质量的碳膜。所要求的设备将简化我们的样品准备程序，并使我们能够有效地为低温EM准备样品。该设备还将用于材料研究中的低温电磁应用，该研究由Gianluigi Botton博士和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共同完成。推进我们在结构生物学方面的研究计划肯定会增加我们制定抗击对人类健康构成重大威胁的疾病的战略的机会。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，最先进的仪器设备转化为一流的培训。所需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最佳的培训经验。

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.