Lucas Miriam Susanna, Schmidt Katarina Wendy, Münz Christian
Scientific Center for Optical and Electron Microscopy (ScopeM), ETH Zurich, 8093 Zurich, Switzerland.
Viral Immunobiology, Institute of Experimental Immunology, University of Zurich, 8057 Zurich, Switzerland.
STAR Protoc. 2025 Aug 14;6(3):104020. doi: 10.1016/j.xpro.2025.104020.
Volume correlative light and electron microscopy (vCLEM) enables ultrastructural analysis of rare cellular events. Here, we present a protocol for targeting virus-containing amphisomes in eukaryotic cells with vCLEM. We describe steps for identifying target cells using confocal fluorescence microscopy, preparing and mounting samples for focused ion beam-scanning electron microscopy (FIB-SEM), relocating the target cells and FIB-SEM stack acquisition, and correlation of volume light and electron microscopy data for 3D modeling. This protocol is optimized for imaging adherent cultured cells. For complete details on the use and execution of this protocol, please refer to Schmidt et al..
体积相关光电子显微镜（vCLEM）能够对罕见的细胞事件进行超微结构分析。在此，我们展示了一种利用vCLEM靶向真核细胞中含病毒双膜体的方案。我们描述了使用共聚焦荧光显微镜识别靶细胞、为聚焦离子束扫描电子显微镜（FIB-SEM）制备和固定样品、重新定位靶细胞并进行FIB-SEM堆栈采集，以及对体积光镜和电镜数据进行关联以进行三维建模的步骤。该方案针对贴壁培养细胞成像进行了优化。有关此方案的使用和执行的完整详细信息，请参考施密特等人的文献。
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