HVEM OPERATION AND IMPROVEMENTS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。摘要：奥尔巴尼大学是世界上仅有的两所致力于生物学研究的大学之一(另一所在日本冈崎)。HVEM的最大加速电压为1200KV，点对点分辨率约为5？它配备了许多专门的样品夹持器，由内部制造，适用于高达75°的单倾斜和双倾斜层析成像。还提供侧向进气式差动泵浦环境室。采用自行设计的增强型视频率CCD电视系统，可以在极低的电子辐照下观察图像，在层析数据采集过程中对图像进行聚焦和跟踪。设想了几个改进领域，以提高清洁发展机制的有用性。其中包括倾斜阶段的计算机控制和低温层析成像的实施。用于电视观看的摄像机的灵敏度足以在冷冻断层扫描过程中进行手动对焦和跟踪，最高可达10,000倍，现有的冷冻传输阶段很可能可以修复。对急速冷冻的整个细胞进行的高压电子显微镜冷冻断层扫描将补充在这类细胞外围的IVEM上所做的工作。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ABSTRACT: The Albany HVEM is one of only two in the world that are dedicated to biological studies (the other is in Okazaki, Japan). The HVEM has a maximum accelerating voltage of 1200kV, and a point-to-point resolution of about 5¿. It is equipped with a number of specialized specimen holders, built in-house, that are suitable for single and double-tilt tomography up to 75¿ tilt. A side-entry differentially-pumped environmental chamber is also available. An in-house designed intensified video-rate CCD TV system can be used to observe the image with very low electron irradiation, for focusing and following the image during tomographic data collection. Several areas of improvement are envisioned for increasing the usefulness of the HVEM. Among these are computer control of the tilt stage and implementation of cryo-tomography. The video camera used for televiewing is sensitive enough for manual focus and tracking during cryo-tomography at up to about 10,000X, and the existing cryotransfer stage can likely be repaired. HVEM cryo-tomography of plunge-frozen whole cells will complement work done on the IVEM on the periphery of such cells.