SEM for serial block-face imaging

摘要 布朗大学请求支持FEI Teneo VS变压扫描电子显微镜 (扫描电子显微镜)，用于生命科学中的连续分块人脸成像。超薄切片在显微镜下切开， 从新切割的块面获取一系列图像，同时丢弃部分。这个 自动化系统能够以各向同性10x10x10 nm分辨率成像大型Z堆栈。一系列的 图像可以用于细胞和组织的3D重建，具有足够的分辨率来识别 小泡、细胞器和神经连接。用户组要求安装基本的Teneo VS系统。 该系统包括对数据进行连续块面成像和三维重建所必需的设备 套装，但不包括其他功能或附件。所要求的系统将取代25年的 布朗大学生物和医学系的老扫描电子显微镜。Teneo VS将是唯一一款 生物医学部和唯一的变压扫描电子显微镜用于连续块面成像 罗德岛州。布朗大学的15名研究人员将立即使用该显微镜 和罗德岛医院进行细胞和亚细胞结构的高分辨率3D成像 从胚胎的种质到视网膜的神经回路。这15个用户总共由26个用户提供资金 美国国立卫生研究院拨款。主要用户对用于连续块面成像的扫描电子显微镜有很大的需求。 作为这一实质性需求的一个迹象，所有主要用户都已获得初步结果，包括以下各项： 用于连续块面成像的状态扫描电子显微镜。虽然这些初步结果表明主要用户 由于集团的需求和专业知识，主要用户对用于连续块脸成像的扫描电子显微镜的访问有限 并且只能想象对他们的研究计划重要的样本的子集。次要用户 无法获得用于连续块脸成像的SEM，但有研究项目将 立即从提议的设备中受益。拟议的成像系统的影响将会扩大 除了主要用户和次要用户的项目，因为显微镜将提供给 罗德岛州更广泛的科学界，有600多个由NIH资助的活跃项目， 年度预算总额为1.84亿美元。该显微镜将安装在Leduc生物成像设施中 布朗大学拥有维护该仪器的技术专长，培训其用户并提供 就已规划和正在进行的研究项目进行咨询。此外，用户还可以访问 协助样品准备和数据分析。Leduc生物成像设施，由 基础广泛的咨询委员会，拥有良好的记录，维护各种高端成像 并拥有强有力的机构支持，这保证了该工具将被用于其 未来几年的最大潜力。

Summary Brown University requests support for a FEI Teneo VS variable pressure Scanning Electron Microscope (SEM) for serial block face imaging in the life sciences. Ultrathin sections are cut inside the microscope and series of images are acquired from the freshly cut block face, while the sections are discarded. The automated system is capable of imaging large z-stacks at isotropic 10x10x10 nm resolution. The series of images can be processed for 3D reconstruction of cells and tissues, with sufficient resolution to identify small vesicles, organelles and neural connections. The user group requested the basic Teneo VS system. This system includes equipment essential for serial block-face imaging and 3D reconstruction of the data sets, but does not include additional features or accessories. The requested system will replace a 25-year old SEM in the Division of Biology and Medicine at Brown University. The Teneo VS will be the only SEM in the Division of Biology and Medicine and the only variable pressure SEM for serial block-face imaging in the State of Rhode Island. The microscope will be used immediately by 15 investigators at Brown University and Rhode Island Hospital for high-resolution 3D imaging of cellular and sub-cellular structures ranging from germ plasm in embryos to neural circuits in the retina. Collectively, these 15 users are funded by 26 NIH grants. The Major Users have a substantial need for the requested SEM for serial block-face imaging. As an indication of this substantial need, all Major Users have obtained preliminary results with various out- of-state SEMs for serial block-face imaging. While these preliminary results demonstrate the Major User group's need and expertise, the major users had limited access to the SEMs for serial block-face imaging and could only image a subset of samples that were important to their research programs. The Minor Users were unable to gain access to SEMs for serial block-face imaging, but have research projects that would immediately benefit from the proposed equipment. The impact of the proposed imaging system will extend beyond the projects of the Major and Minor Users, since the microscope will be made be made available to the broader scientific community in Rhode Island, which has more than 600 active NIH-funded projects with a total annual budget of $184 million. The microscope will be set up in the Leduc Bioimaging Facility at Brown University, which has the technical expertise to maintain the instrument, train its users and provide consultation on planned and ongoing research projects. In addition, the users will have access to assistance in sample preparation and data analysis. The Leduc Bioimaging Facility, supported by a broadly-based Advisory Committee, has an excellent track-record maintaining various high-end imaging systems and has strong institutional support, which guarantees that the instrumentation will be used to its maximum potential for years to come.