Request for a ThermoFisher Helios 5UC DualBeam

7.项目摘要/摘要： 本申请是为购买ThermoFisher Helios 5UC双束聚焦离子束扫描仪提供资金 电子显微镜(FIB-SEM)系统。该仪器是由NIMH资助的一组人要求的 非常需要获取三维(3D)体积的脑组织成像的研究人员 纳米分辨率。所要求的FIB-SEM系统将安装在 匹兹堡大学医学院(UPSOM)的翻译神经科学计划(TNP)。 安装所需的Helios 5 UC双束FIB-SEM是对现有设备的重要扩展 共享的显微镜设备，专门用于脑组织成像。 对3D大体积电子显微镜(VEM)方法的迫切和普遍的需求允许 询问复杂行为及其干扰的大脑机制的基础的研究 精神障碍。3D体电子显微镜(VEM)的出现通过以下方式满足了这一关键需求 生成单独解析的突触和突触亚细胞器的非常详细的连接图谱 在密集堆积的大量脑组织中。所请求的FIB-SEM系统允许自动 以及脑组织的定量VEM成像，避免了与其他VEM相关的许多限制 接近了。尽管FIB-SEM在神经科学研究中的应用相对较新，但我们证明了这一点 技术可以应用于死后的人脑组织，以创建体积的3D、信息丰富的数据集 准备好进行定量分析的脑组织神经纤维。因此，有了这项技术，我们可以审问罚款 神经系统的结构，并准确地找出导致大脑功能障碍的结构紊乱。 NIMH资助的研究人员的巨大反响也证明了对这种仪器的需求 在UPSOM中。我们清楚地解释了所要求的FIB-SEM将如何推进当前的研究目标 NIMH资助的项目，以及这一资源如何对产生新的NIMH赠款申请具有宝贵价值。这个 这项提案中的调查人员在传统EM方法方面有数十年的经验，但这些方法 无法生成由FIB-SEM成像生成的数量或质量的数据。因此，有一个单一的 此应用程序的主要原因：提供急需的、尖端的、高质量的自动化VEM成像 提供给UPSOM内支持NIMH的用户。

7. Project Summary/Abstract: This request is for funds to purchase a ThermoFisher Helios 5UC DualBeam Focused Ion Beam Scanning Electron Microscope (FIB-SEM) system. This instrument is being requested by a group of NIMH-funded investigators who have a significant need to acquire 3-dimensional (3D) volumes of brain tissue imaged at nanometer resolution. The requested FIB-SEM system will be housed in a dedicated facility within the Translational Neuroscience Program (TNP) at the University of Pittsburgh School of Medicine (UPSOM). Installation of the requested Helios 5 UC DualBeam FIB-SEM represents an important expansion of existing shared microscopy facilities that are specialized for, and dedicated to, brain tissue imaging. There is a pressing and prevalent need for 3D large volume electron microscopy (VEM) approaches that permit studies interrogating the basis of brain mechanisms underlying complex behaviors and their disruptions in psychiatric disorders. The advent of 3D volume electron microscopy (VEM) addresses this critical need by generating highly detailed connectomic maps of individually resolved synapses, and the sub-synaptic organelles within, across densely packed, large volumes of brain tissue. The requested FIB-SEM system permits automated and quantitative VEM imaging of brain tissue that avoids many of the limitations associated with other VEM approaches. Although FIB-SEM application to neuroscience research is relatively recent, we show that this technology can be applied to postmortem human brain tissue to create 3D, information-rich datasets of volumes of brain tissue neuropil ready for quantitative analysis. Thus, with this technology we can interrogate the fine structure of the nervous system and pinpoint structural disruptions contributing to brain dysfunction. The need for this instrument is also demonstrated by the overwhelming response from NIMH-funded researchers in the UPSOM. We clearly explain how the requested FIB-SEM will advance the research objectives of current NIMH-funded projects and how this resource is invaluable for generating new NIMH grant applications. The investigators in this proposal have decades of experience in traditional EM approaches, but these methods cannot generate the quantity, or the quality, of data generated by FIB-SEM imaging. Therefore, there is a single prime reason for this application: to provide critically needed, cutting-edge, high quality automated VEM imaging to NIMH supported users within the UPSOM.