Kievits Arent J, Duinkerken B H Peter, Lane Ryan, de Heus Cecilia, van Beijeren Bergen En Henegouwen Daan, Höppener Tibbe, Wolters Anouk H G, Liv Nalan, Giepmans Ben N G, Hoogenboom Jacob P
Department of Imaging Physics, Delft University of Technology, Delft, The Netherlands.
Department of Biomedical Sciences, University Medical Center Groningen, Groningen, The Netherlands.
Methods Microsc. 2024 Jul 11;1(1):49-64. doi: 10.1515/mim-2024-0005. eCollection 2024 Apr.
Elucidating the 3D nanoscale structure of tissues and cells is essential for understanding the complexity of biological processes. Electron microscopy (EM) offers the resolution needed for reliable interpretation, but the limited throughput of electron microscopes has hindered its ability to effectively image large volumes. We report a workflow for volume EM with FAST-EM, a novel multibeam scanning transmission electron microscope that speeds up acquisition by scanning the sample in parallel with 64 electron beams. FAST-EM makes use of optical detection to separate the signals of the individual beams. The acquisition and 3D reconstruction of ultrastructural data from multiple biological samples is demonstrated. The results show that the workflow is capable of producing large reconstructed volumes with high resolution and contrast to address biological research questions within feasible acquisition time frames.
阐明组织和细胞的三维纳米级结构对于理解生物过程的复杂性至关重要。电子显微镜（EM）提供了可靠解释所需的分辨率，但电子显微镜有限的通量阻碍了其对大体积样本进行有效成像的能力。我们报告了一种使用FAST-EM进行体积电子显微镜成像的工作流程，FAST-EM是一种新型的多束扫描透射电子显微镜，通过与64个电子束并行扫描样本，加快了采集速度。FAST-EM利用光学检测来分离各个电子束的信号。展示了从多个生物样本中获取超微结构数据并进行三维重建的过程。结果表明，该工作流程能够在可行的采集时间范围内，以高分辨率和高对比度生成大型重建体积，以解决生物学研究问题。
