Spinning Disk Super Resolution Confocal Microscope for the Vanderbilt Brain Institute

项目摘要/摘要 需要资金购买尼康旋转磁盘超分辨率(CSU-W1/SORA)Conocus 范德比尔特脑研究所内专注于精神健康研究的成像项目的显微镜 (VBI)，一个由范德比尔特大学(VU)和范德比尔特大学(Vanderbilt)教职员工组成的多学科研究机构 大学医学中心(VUMC)。所请求的仪器将整合到 Vanderbilt细胞成像共享资源(CISR)，一个拥有20多年成功历史的显微镜核心 显微镜管理、培训和新技术的获取。这台可共焦的仪器 实时样品中的成像和超分辨率成像是VBI内6个主要用户和7个次要用户的理想选择 围绕突触发生、突触组装和可塑性、突触下信号转导以及 细胞和组织群体之间的交流。这些分子和细胞研究将会取得进展 在2012财年，NIMH和NIH已经在社交、冲动领域资助了总计超过680万美元的项目 攻击性、学习和记忆、Rett综合征(RTT)、ADHD等。 建议的显微镜配置基于VBI和主要用户对实时成像的需求 细胞和亚细胞分辨率。尼康CSU-W1索拉共聚焦显微镜包括：1)CSU-W1 旋转盘加上横河苍井空超分辨率旋转盘，XY超分辨率接近120 nm；2) 3)Z-Piezo阶段，用于提高Z中3D的速度 体积；4)每秒30-100帧的滨松Orca-Fusion sCMOS摄像机；5)激励激光 405、488、561和633 nm；以及6)用于活细胞成像的高分辨率物镜，范围从10x，20x， 40x、60x和100x。这一新系统将设在医学研究大楼III的现有中央科学研究中心空间内 (MRBIII)VBI总部和大多数主要用户实验室所在的地方。 范德比尔特大学正在提供资金支持，以促进该工具的长期运行。如果 基础科学院长办公室承诺提供75,000美元用于支付《公约》工作人员的工资， 购买和升级计算机，并在前5年降低用户费用。技术支持和 新用户的培训将由具有20多年经验的《销售公约》中的杰出工作人员提供便利 在校园内协助300多个实验室的历史。此外，这种设备将成为范德比尔特尼康的一部分 位于《公约》内的卓越中心(COE)。该CIR/尼康COE提供专门的本地 专家团队，支持校园内研究人员的显微镜需求。总的来说，建议的 协调中心系统，加上出色的机构和核心设施支助，将使长期 目的：支持和推进神经科学和心理健康研究。

PROJECT SUMMARY / ABSTRACT Funds are requested to purchase a Nikon Spinning Disk Super Resolution (CSU-W1/SoRa) Confocal microscope for imaging projects focused around mental health research within the Vanderbilt Brain Institute (VBI), a multi-disciplinary research institute comprised of faculty at Vanderbilt University (VU) and Vanderbilt University Medical Center (VUMC). The requested instrument will be integrated into and managed by the Vanderbilt Cell Imaging Shared Resource (CISR), a microscopy core with a 20+ year history of successful microscopy management, training, and acquisition of new technology. This instrument capable of confocal imaging and super resolution imaging in live samples is ideal for 6 Major and 7 Minor Users within the VBI with projects surrounding synaptogenesis, synapse assembly and plasticity, subsynaptic signaling, and communication between cell and tissue populations. These molecular and cellular investigations will advance already funded NIMH and NIH projects totaling over $6.8M in FY22 in the areas of social interaction, impulsive aggression, learning and memory, Rett syndrome (RTT), ADHD, and more. The proposed microscope configuration is based on the needs of the VBI and Major Users for live imaging at cellular and subcellular resolution. The Nikon CSU-W1 SoRa confocal microscope includes: 1) CSU-W1 spinning disk plus Yokogawa SoRa super resolution spinning disk for super resolution near 120 nm in XY; 2) stage-top incubation for imaging live cells and organoids; 3) Z-Piezo stage for increased speed in Z for 3D volumes; 4) Hamamatsu Orca-Fusion sCMOS camera with 30-100 frames per second; 5) excitation lasers of 405, 488, 561, and 633 nm; and 6) objective lenses for live cell imaging at high resolution ranging from 10x, 20x, 40x, 60x, and 100x. This new system would be housed in existing CISR space in Medical Research Building III (MRBIII) where the VBI is based and the majority of the Major User labs are located. Vanderbilt University is providing financial support to promote the long-term operation of the instrument. If awarded, the Office of the Dean of Basic Sciences is committed to providing $75,000 for CISR staff salary, computer purchases and upgrades, and reduction in user fees over the first 5 years. Technical support and training of new users will be facilitated by the exceptional staff within the CISR, which has a greater than 20-year history of assisting over 300 labs on campus. Moreover, this equipment will become part of the Vanderbilt Nikon Center of Excellence (COE) that is housed within the CISR. This CISR/Nikon COE provides a dedicated local team of specialists to support the microscopy needs of investigators on campus. Collectively, the proposed confocal system, together with outstanding institutional and core facility support, will enable the long-term objective of supporting and advancing neuroscience and mental health research Vanderbilt.