Integrated ultramicrotome for serial block face scanning electron microscopy (SBF-SEM)

用于串行块面扫描电子显微镜 (SBF-SEM) 的集成超薄切片机

• 批准号: 500508043
• 金额: --
• 依托单位: Charité - Universitätsmedizin Berlin
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/500508043?language=en
• 关键词: Integrated; ultramicrotome; serial; block; face

Recent technical developments in biomedical electron microscopy (EM) aim at visualizing structures (e.g. cells or tissue samples) in their topographic context at high resolution in 3D (volume EM). The analysis of large sample volumes allows to reveal biological "connectomes" in their spatial complexity.We apply for an ultramicrotome, integrated into the column of a scanning electron microscope, for automated 3D analyses by serial block face scanning electron microscopy (SBF-SEM). The system will complement a scanning electron microscope with field emission cathode (FEG-SEM) operated since 2019 by the Core Facility Electron Microscopy (CFEM) of Charité - Universitätsmedizin Berlin. The ultramicrotome is mounted on a table and can be integrated via the front of the FEG-SEM column. By exchange of the table, the FEG-SEM can either be operated in conventional mode or in SBF-SEM mode.An essential approach for generating 3D image datasets in SEM is based on the principle of mechanical removal of thin slices from the sample surface, alternating to imaging the sample surface after each removal. In SBF-SEM, this removal from the sample surface is realized by the principle of ultramicrotomy (i.e. with a diamond knife). After proper alignment, the acquisition of raw data runs automatically. The maximum spatial resolution is about 10-15 (x/y) and 25-50 nm (z), respectively. The maximum sample volume is in the order of about 1 mm (z) over an area of about 0.5 x 0.5 mm (x/y).The microscope, upgraded for SBF-SEM investigations, will be made available to all interested users according to the user policy of the CFEM of Charité - Universitätsmedizin Berlin. Furthermore, it will be embedded into the research infrastructure of the Berlin University Alliance (BUA).

生物医学电子显微镜 (EM) 的最新技术发展旨在以 3D 高分辨率可视化结构（例如细胞或组织样本）（体积 EM），对大样本体积的分析可以揭示其结构中的生物“连接组”。空间复杂性。我们申请将超薄切片机集成到扫描电子显微镜镜筒中，通过串行块面扫描电子显微镜 (SBF-SEM) 进行自动 3D 分析。该系统将补充扫描电子显微镜。带有场发射阴极的显微镜 (FEG-SEM) 自 2019 年起由柏林夏里特医学大学的核心设施电子显微镜 (CFEM) 运行。超薄切片机安装在工作台上，可通过 FEG-SEM 柱的前端进行集成。通过交换表格，FEG-SEM 可以在传统模式或 SBF-SEM 模式下运行。在 SEM 中生成 3D 图像数据集的基本方法基于从样品表面机械去除薄片的原理，在每次去除后交替对样品表面进行成像，在 SBF-SEM 中，这种从样品表面的去除是通过超薄切片原理（即使用金刚石刀）实现的。正确对齐后，原始数据的采集会自动运行，最大空间分辨率分别约为 10-15 nm (x/y) 和 25-50 nm (z)，最大样品体积约为 1 mm (z)。 ）面积约为 0.5 x 0.5 毫米 (x/y)。根据柏林夏里特大学 CFEM 的用户政策，为 SBF-SEM 研究升级的显微镜将提供给所有感兴趣的用户。此外，它将被嵌入到柏林大学联盟（BUA）的研究基础设施中。