CryoEM Equipment Enhancements for Florida State University

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）是佛罗里达州低温电子显微镜和3-D电子显微镜的焦点。没有其他大学的设施有类似的重点或类似的设施。本申请请求资金，以减轻对专用于制备薄玻璃体冰膜的设备和专用于电子断层扫描的标本持有人的迫切需求。该设备将支持主要的BSIR用户，博士的研究。Kenneth Taylor， Kenneth Roux和Thomas Roberts以及来自长期合作伙伴Michael Chapman博士实验室的访客，他最近搬到了俄勒冈健康与科学大学。申请资金用于4项主要设备，包括(1)Vitrobot计算机控制的用于制备玻璃化悬浮液的样品冷冻机，(2)用于制备亲水多孔碳膜的等离子体清洁器，(3)高倾斜低温电子断层扫描样品架和(4)双轴高倾斜室温样品架。薄玻璃体冷冻水合悬浮液的研究是FSU冷冻电镜研究重点的很大一部分，但这些标本的制备目前是许多项目的瓶颈。通过购买用于制作亲水支撑膜的组合等离子清洁剂和用于制备玻璃化薄水膜的Vitrobot，可以消除这一瓶颈。电子断层扫描是三个申请小组的主要重点，而第四个小组正在开始探索其可能性。双轴断层扫描支架将提高快速冷冻、冷冻替代、塑料嵌入生物标本重建的通量和质量，而低温断层扫描支架将增强冷冻水合标本的三维成像。这些项目将支持NIH资助的以下研究：(1)肌原性肌肉产生力的基本机制，这与心肌功能的健康相关；(2)平滑肌肌球蛋白调节，与高血压健康相关；(3)与癌症健康相关的粘附结构和迁移细胞；(4)艾滋病毒和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。

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. No other University facility has a comparable emphasis or comparable facilities. This application requests funds to alleviate a critical need for equipment dedicated to the preparation of thin vitreous ice films and for specimen holders dedicated to electron tomography. This equipment will support the research of the major BSIR users, Drs. Kenneth Taylor, Kenneth Roux and Thomas Roberts and visitors from the lab of a long standing collaborator, Dr. Michael Chapman, who recently moved to Oregon Health & Science University. Funds are requested for 4 main items of equipment including (1) Vitrobot computer controlled specimen freezer for preparing thin vitrified suspensions, (2) a plasma cleaner for producing hydrophilic holey carbon films, (3) a high tilt cryoelectron tomography specimen holder and (4) a dual axis, high tilt, room temperature specimen holder. The study of thin, vitreous, frozen hydrated suspensions is a large part of the cryoEM research emphasis at FSU, but preparation of these specimens is currently a bottleneck in a number of projects. This bottleneck will be removed by purchase of the combination plasma cleaner for making hydrophilic support films and Vitrobot for preparing the thin vitrified aqueous films. Electron tomography is a major emphasis of three applicant groups while the fourth is beginning to explore the possibilities. The dual axis tomography holder will improve both throughput and quality of reconstructions obtained from rapidly frozen, freeze substituted, plastic embedded biological specimens while the cryotomography holder will enhance 3-D imaging of frozen hydrated specimens. These items 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) adhesion structures and migrating cells, which has health relevence in cancer; (4) the structure of HIV and SIV viruses which are fundamental to understanding AIDS; and (5) the development of Adeno Associated Virus as a gene therapy vector.