Acquisition of Detector for JEOL 3200 FSC Electron Microscope

采购 JEOL 3200 FSC 电子显微镜检测器

• 批准号: 8825827
• 负责人: BRIDGET Olivia CARRAGHER
• 金额: $ 60万
• 依托单位: NEW YORK STRUCTURAL BIOLOGY CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8825827
• 关键词: Adopted; Bacteriophages; Cell Adhesion; Cell physiology; Communities; Complex; Contract Services; Crystallography; Development; Doctor of Philosophy; Educational workshop; Electron Microscope; Electron Microscopy; Electrons; Film; Freezing; Genetic Transcription; Helium; Hepatitis C virus; Housing; Image; Imaging Device; Imaging technology; Institution; Laboratories; Liquid substance; Maintenance; Malignant Neoplasms; Membrane Proteins; Metabolic Diseases; Microscope; Microsporidia; Neurodegenerative Disorders; New York; Newsletter; Nitrogen; Nuclear; Positioning Attribute; Preparation; Resolution; Resource Sharing; Resources; Roentgen Rays; Sampling; Source; Specimen; Staging; Structure; Synchrotrons; System; Technology; Temperature; Time; Translations; Wages; base; charge coupled device camera; detector; electron crystallography; experience; image processing; imaging detector; improved; instrument; instrumentation; interest; macromolecule; new technology; operation; outreach; particle; pathogen; structural biology; tomography

 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.

 描述（由申请人提供）：纽约结构生物学中心 (NYSBC) 是由纽约九个学术机构共享的仪器资源，拥有冷冻电子显微镜 (cryo-EM)、核磁共振波谱仪和包括光束在内的 X 射线设施。通过这种方式，NYSBC 提供了整个结构生物学领域的高端资源，以维持其尖端设施。电子显微镜方面，我们正在寻求升级高端冷冻电镜上的成像探测器。具体来说，我们的 JEOL 3200FSC 电子显微镜采用 300 keV 场发射源，配有柱内能量过滤器和基于墨盒的样品台，可提供出色的性能。在液氮或液氦温度下的稳定性 在 2005 年购买时，最先进的成像设备是 4k x 4k CCD 相机，目前用于各种项目单粒子、断层扫描和二维晶体学然而，这种成像技术对我们的用户的高分辨率结构测定产生了限制。在过去的几年中，直接电子探测器取得了快速发展，并且已被证明优于这两种技术。 CCD 相机和胶片。特别是 Gatan K2 Summit 相机已被许多实验室采用，并在生物学上重要的大分子方面产生了显着的结果，这些结果激起了人们对使用冷冻电镜进行结构测定的新兴趣。通过为我们的 JEOL 3200FSC 显微镜配备这款相机，我们的目标是使 NYSBC 能够为我们不同的用户群体提供这些新机会。特别是，此次升级将显着提高图像质量，从而提高当前项目的分辨率并促进新项目的发展。该应用中描述的项目包括对丙型肝炎病毒、微孢子虫和噬菌体等未染色冷冻样本进行断层扫描，以及细胞中涉及的复合物的单粒子分析。这些项目将对我们对侵入性病原体以及与癌症、代谢和神经退行性疾病相关的重要细胞过程的理解产生重大影响。作为 PI，NYSBC EM 设施拥有三名全职博士级员工，他们在样品制备、EM 操作和计算图像处理方面提供了丰富的实践经验。通过 NYSBC 每年提供的为期一个学期的课程来实现对社区的宣传，同时管理定期的高级研讨会以及向我们不断扩大的 NYSBC 社区分发的季度通讯，通过支持员工工资和服务来提供强有力的机构承诺。充分利用这项令人兴奋的新技术所需的所有电子显微镜和计算设施的合同。