High-resolution and high-performance CCD camera for transmission electron microscopy

我正在使用冷冻电子显微镜和图像分析技术来研究大分子复合体如何在细胞中执行特定功能，特别是了解蛋白质跨细胞膜运输的机制。在纳米级详细检查蛋白质和其他生物分子等细胞成分的能力，对于了解细胞在健康和疾病中的运作方式至关重要。为了进行研究，我接触到了麦吉尔大学最先进的冷冻扫描透射式电子显微镜(CRYO-S/TEMS)、FEI Tecnai G2 F20 200千伏CRYO-S/TEM机和FEI TITAN KRIOS 300 KVCRYO-S/TEM机。在这些显微镜下对标本的研究是劳动密集型和昂贵的，因此，我使用FEI Tecnai 12 120KV透射电子显微镜(‘T12’)来获得通过负染和铂-C复制品制备的蛋白质和细胞器结构的高分辨率图像。然而，T12配备了一个低分辨率的100万像素的ccd摄像头，不足以为我的标本成像。目前的应用是申请资金，为T12配备现代、高分辨率和高性能的800万像素CCD相机，使我能够记录样品中存在的全部信息，以便确定分离的大分子组件的三维结构。新的CCD摄像机将显着提高数据收集的效率，并改善高素质人员和研究人员的培训机会。

I am using techniques of cryo-electron microscopy (cryoEM) and image analysis to investigate how macromolecular complexes perform specific functions in cells, and in particular, to understand the mechanisms of protein transport across cellular membranes. The ability to examine cellular components like proteins and other biological molecules at nano-scale detail is crucial to understanding how cells operate in health and disease. For my research, I have access to state-of-the-art cryo-scanning-transmission electron microscopes (cryo-S/TEMs) at McGill University; the FEI Tecnai G2 F20 200 kV cryo-S/TEM and the FEI Titan Krios 300 kV cryo-S/TEM acquired through a recent Canada Foundation for Innovation Grant. The investigation of specimens in these microscopes is labour intensive and expensive, therefore, I use the FEI Tecnai 12 120 kV TEM ('T12') to obtain high-resolution images of protein and organelle structures prepared by negative staining and Pt-C replicas. The T12 is, however, equipped with a low-resolution 1 megapixel CCD camera that is insufficient for the imaging of my specimens. The current application is to request funds to equip the T12 with a modern, high-resolution and high-performance 8 megapixel CCD camera that will enable me to record the full information present in the samples in order to determine the 3-D structures of isolated macromolecular assemblies. The new CCD camera will significantly increase the efficiency of data collection and improve training opportunities for highly qualified personnel and researchers.