Purchase of a FEI Titan Krios for 3-D EM

购买用于 3-D EM 的 FEI Titan Krios

• 批准号: 7498219
• 负责人: KENNETH ALLEN TAYLOR
• 金额: $ 200万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7498219
• 关键词: 15 year old; 3-Dimensional; Acquired Immunodeficiency Syndrome; Age; Anderson syndrome; Arts; Biological; Biological Sciences; Computer software; Cryoelectron Microscopy; Data; Defect; Dependovirus; Development; Electron Microscope; Electron Microscopy; Faculty; Florida; Freezing; Funding; Gene Transduction Agent; Generations; HIV; Health; Health Sciences; Hypertension; Image; Malignant Neoplasms; Methods; Microscope; Minor; Muscle; Muscle function; Myocardium; Numbers; Oregon; Phase; Plastic Embedding; Regulation; Request for Applications; Research; Resources; Robotics; Running; SIV; Series; Smooth Muscle Myosins; Specimen; Structure; Support of Research; Suspension substance; Suspensions; Thick; Time; Titan; United States National Institutes of Health; Universities; Vesicle; Virus; Work; cell motility; electron tomography; improved; instrument; instrumentation; member; particle; reconstruction; software development; tomography; trafficking

DESCRIPTION (provided by applicant): 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 center piece of this facility, an FEI CM 300-FEG electron microscope with a Titan-Krios, a robotic electron microscope with enhanced capabilities for cryoEM and electron tomography. Our CM 300-FEG microscope is 15 years old and is showing its age both in terms of down time and our ability to implement instrumentation and software developments produced by others in the field. The instrument lacks an imaging filter which is crucial for work that now involves thick specimens, and will become increasing problematic for retrofit of new developments in electron microscopy such as phase plates and aberration correctors. The requested Titan-Krios will support the research of the major BSIR users, Drs. Kenneth Taylor, Kenneth Roux and Thomas Roberts, new faculty member Dr. Scott Stagg and visitors from the lab of a long standing collaborator, Dr. Michael Chapman, who recently moved to Oregon Health & Science University. Three minor projects are described that will benefit from access to a modern electron microscope. The Titan-Krios will also be able to support remote users, making it possible to offer its extensive capabilities conveniently to users outside of the university and become a true research resource for the Southeast. The study of thin, vitreous, frozen hydrated suspensions, both using electron tomography and single particle methods is a large and growing cryoEM research emphasis at FSU, but obtaining sufficient data, either as tilt series or projections is currently a bottleneck in a number of projects. The robotic capabilities of the Titan-Krios as well as the ability to run state of the art automated EM software, such as Leginon, will alleviate this bottleneck. Electron tomography is a major emphasis of three applicant groups while the other two are beginning to explore the possibilities. The dual axis tomography capabilities of the Titan-Krios will improve both throughput and quality of reconstructions obtained from both frozen hydrated as well as rapidly frozen, freeze substituted, plastic embedded biological specimens. 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) smooth muscle myosin regulation, which has health relevance in hypertension; (3) structures involved in cell migration, which has health relevence in cancer; (4) the structure of HIV and SIV viruses which are fundamental to understanding AIDS; (5) the development of Adeno Associated Virus as a gene therapy vector and (6) NIH related research into the structure of COPII vesicles, which are implicated in defects of cargo trafficking including chylomicron retention disease and cranio-lenticulo-sutural displasia.

描述（由申请人提供）：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冷冻电子显微镜和 3D 电子显微镜的焦点。没有其他大学设施具有可比的重点或设施。该申请要求资金来更换和升级该设施的核心部件，即配备 Titan-Krios 的 FEI CM 300-FEG 电子显微镜，Titan-Krios 是一款具有增强的冷冻电镜和电子断层扫描功能的机器人电子显微镜。我们的 CM 300-FEG 显微镜已有 15 年的历史，无论是在停机时间还是我们实施该领域其他公司生产的仪器和软件开发的能力方面都显示出它的年龄。该仪器缺乏成像滤光片，这对于现在涉及厚样本的工作至关重要，并且对于电子显微镜新发展（例如相位板和像差校正器）的改造将成为越来越多的问题。所请求的 Titan-Krios 将支持主要 BSIR 用户的研究，博士。肯尼思·泰勒 (Kenneth Taylor)、肯尼思·鲁克斯 (Kenneth Roux) 和托马斯·罗伯茨 (Thomas Roberts)、新任教员斯科特·斯塔格 (Scott Stagg) 博士以及来自长期合作者迈克尔·查普曼 (Michael Chapman) 博士实验室的访客，后者最近搬到了俄勒冈健康与科学大学。描述了三个小型项目，这些项目将受益于现代电子显微镜的使用。 Titan-Krios 还将能够支持远程用户，从而可以方便地向大学以外的用户提供其广泛的功能，并成为东南部真正的研究资源。使用电子断层扫描和单粒子方法对稀薄的玻璃体冷冻水合悬浮液进行研究是 FSU 冷冻电镜研究的一个重要且不断增长的重点，但获取足够的数据（无论是倾斜系列还是投影）目前是许多项目的瓶颈。 Titan-Krios 的机器人功能以及运行最先进的自动化 EM 软件（例如 Leginon）的能力将缓解这一瓶颈。电子断层扫描是三个申请小组的重点，而另外两个申请小组正在开始探索其可能性。 Titan-Krios 的双轴断层扫描功能将提高从冷冻水合以及快速冷冻、冷冻替代、塑料嵌入生物样本获得的重建的吞吐量和质量。 Titan-Krios 将支持 NIH 资助的以下研究：(1) 生肌力产生的基本机制，与心肌功能与健康相关； (2) 平滑肌肌球蛋白调节，与高血压的健康相关； (3) 参与细胞迁移的结构，与癌症的健康相关； (4) HIV 和 SIV 病毒的结构，这是了解艾滋病的基础； (5) 腺相关病毒作为基因治疗载体的开发，以及 (6) NIH 对 COPII 囊泡结构的相关研究，这些囊泡与货物运输的缺陷有关，包括乳糜微粒滞留病和颅-豆状核-缝线发育不良。

项目成果

CryoEM structure of Drosophila flight muscle thick filaments at 7 Å resolution.

果蝇飞行肌粗丝的冷冻电镜结构，分辨率为 7 ×。

• 发表时间: 2020
• 影响因子: 4.4
• 作者: Daneshparvar, Nadia;Taylor, Dianne W;O'Leary, Thomas S;Rahmani, Hamidreza;Abbasiyeganeh, Fatemeh;Previs, Michael J;Taylor, Kenneth A
• 通讯作者: Taylor, Kenneth A

The cryo-EM 3D image reconstruction of isolated Lethocerus indicus Z-discs.

孤立的 Lethocerus indicus Z 盘的冷冻电镜 3D 图像重建。

• 发表时间: 2023-12
• 影响因子: 2.7
• 作者: Yeganeh, Fatemeh Abbasi;Summerill, Corinne;Hu, Zhongjun;Rahmani, Hamidreza;Taylor, Dianne W;Taylor, Kenneth A
• 通讯作者: Taylor, Kenneth A

A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1.

STING 的保守 PLPLRT/SD 基序介导 TBK1 的招募和激活。

• 发表时间: 2019-05
• 影响因子: 64.8
• 作者: Zhao, Baoyu;Du, Fenglei;Xu, Pengbiao;Shu, Chang;Sankaran, Banumathi;Bell, Samantha L;Liu, Mengmeng;Lei, Yuanjiu;Gao, Xinsheng;Fu, Xiaofeng;Zhu, Fanxiu;Liu, Yang;Laganowsky, Arthur;Zheng, Xueyun;Ji, Jun;West, A Phillip;Watson, Robert O
• 通讯作者: Watson, Robert O

Competing scaffolding proteins determine capsid size during mobilization of Staphylococcus aureus pathogenicity islands.

竞争性支架蛋白决定金黄色葡萄球菌致病岛动员过程中衣壳的大小。

• 发表时间: 2017-10-06
• 影响因子: 7.7
• 作者: Dearborn, Altaira D;Wall, Erin A;Kizziah, James L;Klenow, Laura;Parker, Laura K;Manning, Keith A;Spilman, Michael S;Spear, John M;Christie, Gail E;Dokland, Terje
• 通讯作者: Dokland, Terje

The structures of natively assembled clathrin-coated vesicles.

天然组装的网格蛋白包被的囊泡的结构。

• 发表时间: 2020-07
• 影响因子: 13.6
• 作者: Paraan, Mohammadreza;Mendez, Joshua;Sharum, Savanna;Kurtin, Danielle;He, Huan;Stagg, Scott M
• 通讯作者: Stagg, Scott M