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)是佛罗里达州冷冻电子显微镜和3-D电子显微镜的焦点。没有其他大学的设施具有类似的重点或类似的设施。这项申请需要资金，以缓解对专门用于制备薄玻璃冰膜的设备和专门用于电子断层扫描的样品支架的迫切需求。该设备将支持BSIR的主要用户Kenneth Taylor博士、Kenneth Roux博士和Thomas Roberts博士以及来自长期合作伙伴Michael Chapman博士实验室的访客的研究，Michael Chapman博士最近搬到了俄勒冈健康与科学大学。请拨资金用于4项主要设备，包括(1)用于制备薄玻璃悬浮液的Vitbot计算机控制的样品冷冻机，(2)用于生产亲水多孔碳膜的等离子清洁器，(3)高倾斜低温电子断层扫描样品夹具和(4)双轴高倾斜室温样品夹具。薄薄、玻璃质、冷冻水化悬浮液的研究是FSU低温EM研究重点的一大部分，但这些样品的制备目前是许多项目的瓶颈。这一瓶颈将通过购买用于制造亲水支撑膜的组合等离子体清洗器和用于制备薄玻璃化水膜的Vitbot来消除。电子断层扫描是三个申请组的主要重点，而第四个申请组正在开始探索可能性。双轴断层扫描支架将提高从快速冷冻、冷冻替代、塑料嵌入的生物标本中获得的重建的吞吐量和质量，而冷冻断层扫描支架将增强冷冻水合样品的三维成像。这些项目将支持美国国立卫生研究院资助的以下研究：(1)与心脏肌肉功能有关的肌源性力量产生的基本机制；(2)与高血压有关的平滑肌肌球蛋白调节；(3)与癌症有关的黏附结构和迁移细胞；(4)对了解艾滋病至关重要的艾滋病毒和SIV病毒的结构；以及(5)腺相关病毒作为基因治疗载体的发展。

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.