SEM for serial block-face imaging

总结 布朗大学请求支持 FEI Teneo VS 变压扫描电子显微镜 (SEM) 用于生命科学中的连续块面成像。在显微镜内切割超薄切片并 从新切割的块面获取一系列图像，而切片则被丢弃。的 自动化系统能够以各向同性 10x10x10 nm 分辨率对大型 z 堆栈进行成像。该系列的 可以处理图像以进行细胞和组织的 3D 重建，并具有足够的分辨率来识别 小囊泡、细胞器和神经连接。该用户组请求基本的 Teneo VS 系统。 该系统包括串行块面成像和数据 3D 重建所必需的设备 套装，但不包括附加功能或配件。所请求的系统将取代使用了 25 年的系统 布朗大学生物与医学系的老SEM。 Teneo VS 将是唯一的 SEM 生物和医学部门以及唯一用于连续块面成像的可变压力 SEM 罗德岛州。布朗大学的 15 名研究人员将立即使用该显微镜 和罗德岛医院对细胞和亚细胞结构进行高分辨率 3D 成像 从胚胎的种质到视网膜的神经回路。这 15 位用户总共由 26 位用户资助 美国国立卫生研究院拨款。主要用户对用于串行块面成像的 SEM 有着巨大的需求。 作为这一实质性需求的指示，所有主要用户都已获得各种结果的初步结果 - 用于连续块面成像的状态 SEM。虽然这些初步结果表明主要用户 由于小组的需求和专业知识，主要用户对用于串行块面成像的 SEM 的访问有限 并且只能对对其研究项目很重要的一部分样本进行成像。未成年人用户 无法使用 SEM 进行连续块面成像，但有研究项目可以 立即受益于建议的设备。所提议的成像系统的影响将扩大 超出主要和次要用户的项目，因为显微镜将可供 罗德岛州更广泛的科学界，有超过 600 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，位于 布朗大学拥有维护仪器、培训用户并提供 就计划中和正在进行的研究项目进行咨询。此外，用户将有权访问 协助样品制备和数据分析。 Leduc 生物成像设施，由 基础广泛的咨询委员会，在维护各种高端成像方面拥有出色的记录 具有完善的制度体系和强有力的制度支撑，保证了仪器仪表的发挥作用 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.