Acquisition of Detector for JEOL 3200 FSC Electron Microscope

 描述（由申请人提供）：纽约结构生物学中心（NYSBC）是由九个纽约学术机构共享的仪器资源。NYSBC拥有低温电子显微镜（cryo-EM），NMR光谱仪和X射线设备，包括布鲁克海文国家实验室NSLS同步加速器的光束线。通过这种方式，NYSBC在整个结构生物学领域提供高端资源。为了保持电子显微镜的尖端设施，我们正在寻求升级我们的高端冷冻EM上的成像探测器。具体来说，我们的JEOL 3200 FSC电子显微镜具有300 keV场发射源，带有柱内能量过滤器和基于卡盘的样品台，可在液氮或液氦温度下提供出色的稳定性。在2005年购买时，最先进的成像设备是4k x 4k CCD相机，目前用于涉及单粒子，断层扫描和2D晶体学的各种项目。然而，这种成像技术对我们的用户进行高分辨率结构测定产生了限制。在过去的几年中，直接电子探测器得到了迅速的发展，并已被证明大大优于这些CCD相机和胶片。具体来说，Gatan K2 Summit相机已被许多实验室采用，并在生物重要的大分子上产生了引人注目的结果。这些结果引发了新的兴趣，使用冷冻EM结构测定。通过为我们的JEOL 3200 FSC显微镜配备这款相机，我们的目标是使NYSBC能够为我们多样化的用户群体提供这些新的机会。特别是，这一升级将大大改善图像质量，从而提高当前项目的分辨率，并促进现有CCD技术无法实现的新项目。本申请中描述的项目包括未染色冷冻标本的断层扫描，如丙型肝炎病毒，微孢子虫和噬菌体，涉及细胞粘附，转录，翻译，核和细胞质运输的复合物的单颗粒分析，以及各种膜蛋白的电子晶体学。这些项目将对我们了解侵入性病原体以及与癌症、代谢和神经退行性疾病相关的基本细胞过程产生重大影响。除了PI外，NYSBC EM工厂还有三名全职博士，我们拥有一批高水平的工作人员，他们在样品制备、EM操作和计算机图像处理方面具有丰富的实践经验。NYSBC EM设施开发了一个强大的系统，用于维护和管理其仪器的共享使用。外联到社区是通过一个学期的课程，每年在NYSBC提供，虽然定期高级研讨会，并通过季度通讯分发给我们不断扩大的EM社区完成。NYSBC提供了强有力的机构承诺，支持员工工资，所有电子显微镜和计算设施的服务合同，以充分利用这一令人兴奋的新技术。

 DESCRIPTION (provided by applicant): The New York Structural Biology Center (NYSBC) is an instrumentation resource shared by nine New York academic institutions. NYSBC houses cryo-electron microscopes (cryo-EM), NMR spectrometers and X-ray facilities including a beam line at the NSLS synchrotron at Brookhaven National Laboratory. In this way, NYSBC provides high-end resources across the entire spectrum of structural biology. In order to maintain cutting-edge facilities in electron microscopy, we are seeking to upgrade the imaging detector on our top-end cryo-EM. Specifically, our JEOL 3200FSC electron microscope features a 300 keV field emission source with an in-column energy filter and a cartridge based specimen stage that provides excellent stability at either liquid nitrogen or liquid helium temperatures. At the time o purchase in 2005, the state of the art imaging device was a 4k x 4k CCD camera, which is currently used for a variety of projects involving single particles, tomography and 2D crystallography. However, this imaging technology produces a limitation to high resolution structure determination by our users. Rapid developments in direct electron detectors have occurred over the last several years and have been demonstrated to greatly outperform both these CCD cameras and film. Specifically the Gatan K2 Summit camera has been adopted by a number of laboratories and has produced dramatic results on biologically important macromolecules. These results have sparked a renewed interest in using cryo-EM for structure determination. By equipping our JEOL 3200FSC microscope with this camera, we aim to position NYSBC to provide these new opportunities to our diverse group of users. In particular, this upgrade will significantly improve image quality, thereby increasing the resolution for curren projects and facilitate new projects that were impractical with the existing CCD technology. Projects described in this application include tomography of unstained frozen specimens such as hepatitis C virus, microsporidia and bacteriophages, single particle analysis of complexes involved in cell adhesion, transcription, translation, nuclear and cytoplasmic transport, and electron crystallography of various membrane proteins. These projects will have a significant impact on our understanding of invasive pathogens and of essential cellular processes associated with cancer and with metabolic and neurodegenerative diseases. In addition to the PI, the NYSBC EM facility has three full-time, Ph.D. level staff who provide extensive hands-on experience in sample preparation, EM operation and computational image processing. The NYSBC EM facility has developed a robust system for maintenance and for managing the shared use of its instruments. Outreach to the community is accomplished through a semester-long course that is offered annually at NYSBC, though periodic advanced workshops and through a quarterly newsletter that is distributed to our expanding EM community. NYSBC provides a strong institutional commitment by supporting staff salaries, service contracts on all the electron microscopes and computational facilities required to take full advantage of this exciting new technology.