Purchase of a direct electron camera for the Titan-Krios at FSU

为 FSU 的 Titan-Krios 购买一台直接电子相机

基本信息

批准号：
8640487
负责人：
KENNETH ALLEN TAYLOR
金额：
$ 38.37万
依托单位：
FLORIDA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-09 至 2016-09-08
项目状态：
已结题

项目摘要

6 PROJECT SUMMARY/ABSTRACT The Biological Science Imaging Resource (BSIR) at Florida State University (FSU) is the focal point for cryoelectron microscopy and 3-D electron microscopy in the state of Florida and the Southeast. No other University facility has a comparable emphasis or facilities. This application requests funds to replace and upgrade the electron camera on the center piece of this facility, an FEI Titan-Krios electron microscope with a direct electron camera, a revolutionary camera that is far superior to any previous image recording method. We will replace an existing Gatan Ultrascan camera with a Gatan K2 Summit camera. The requested K2 Summit will support the research of the major BSIR users, Drs. Kenneth Taylor, Scott Stagg, Hong Li, and new faculty member Dr. Elizabeth Stroupe. It will also provide visitors to the BSIR from outside FSU access to a state of the art facility. Two new investigators outside of FSU have provided projects requiring access to a modern electron microscope with a direct electron camera. The study of thin, vitreous, frozen hydrated suspensions, using both electron tomography and single particle methods is a large and growing cryoEM research emphasis worldwide. The robotic capabilities of the Titan-Krios with its ability to run state of the art automated EM software, such as Leginon, are well adapted to this need, but ultimately, the quality of the recording medium limits the quality of the final result. The poor MTF of today's CCD cameras is offset somewhat by recording images at higher magnification, but this reduces the field of view requiring even more images. Beam induced specimen motion is recognized as a fundamental problem that can be solved by rapid exposure times, known as movie mode, and local (or global) corrections for particle motion. Movie mode images will be crucial for electron tomography, which relies on images taken at high tilt angles for best results. Such images are prone to beam induced specimen motion perpendicular to the specimen plane, a problem that would be reduced by correcting for motion in 25 msec image recording times. The Titan-Krios will support NIH funded research in (1) the fundamental mechanism of force generation in a myogenic muscle, which has health relevence to cardiac muscle function; (2) myosin motor activity, which has health relevance in multiple diseases; (3) transformation of bacteria by bacteriophage which leads to multidrug resistance; (4) the structure of COPII vesicles, which are implicated in defects of cargo trafficking including chylomicron retention disease and cranio-lenticulo-sutural displasia; (5) The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) which confer adaptive immunity of bacteria against invasive genetic elements and (6) NIH related research into the structure and function of sulfite reductases.

6 项目概要/摘要佛罗里达州立大学 (FSU) 的生物科学成像资源 (BSIR) 是佛罗里达州和东南部的冷冻电子显微镜和 3D 电子显微镜。没有其他大学设施具有类似的重点或设施。此申请要求资金来更换和升级该设施中心部分的电子相机，即一台 FEI Titan-Krios 电子显微镜，配有直接电子相机，一种革命性的相机，远远优于以前的任何图像记录方法。我们将用 Gatan K2 Summit 相机取代现有的 Gatan Ultrascan 相机。所要求的 K2 峰会将支持主要 BSIR 用户的研究，博士。肯尼思·泰勒、斯科特·斯塔格、李红和新教师成员伊丽莎白·斯特鲁普博士。它还将为来自 FSU 外部的 BSIR 访问者提供访问状态的信息艺术设施。佛罗里达州立大学之外的两名新研究人员提供了需要使用现代技术的项目带有直接电子相机的电子显微镜。稀薄的玻璃质冷冻水合悬浮液的研究，使用电子断层扫描和单粒子方法是冷冻电镜研究的一个重要且不断增长的重点全世界。 Titan-Krios 的机器人功能及其运行最先进的自动化 EM 的能力 Leginon 等软件很好地适应了这种需求，但最终取决于记录介质的质量限制了最终结果的质量。当今 CCD 摄像机较差的 MTF 通过记录在一定程度上得到了弥补更高放大倍率的图像，但这会减少视野，需要更多图像。光束感应样本运动被认为是一个基本问题，可以通过快速曝光时间来解决，已知作为电影模式，以及粒子运动的局部（或全局）校正。电影模式图像对于电子断层扫描，依靠以高倾斜角度拍摄的图像来获得最佳结果。此类图像很容易出现对于光束引起的垂直于样本平面的样本运动，这个问题可以通过以下方式减少校正 25 毫秒图像记录时间内的运动。 Titan-Krios 将支持 NIH 资助的研究 (1) 肌原肌产生力的基本机制，与心脏健康相关肌肉功能； (2) 肌球蛋白运动活动，与多种疾病的健康相关； (3)改造噬菌体杀死细菌，导致多重耐药性； (4) COPII囊泡的结构，它们是与货物运输缺陷有关，包括乳糜微粒滞留病和颅骨豆状缝发育不良； (5) 成簇的规则间隔的短回文重复序列 (CRISPR) 细菌针对侵入遗传元件的适应性免疫及（6）NIH相关结构研究和亚硫酸还原酶的功能。