Scanning Electron Microscopy (SEM) and Immuno-SEM of Nuclear Pore Complexes from Amphibian Oocytes, Mammalian Cell Cultures, Yeast, and Plants.
Scanning Electron Microscopy (SEM) and Immuno-SEM of Nuclear Pore Complexes from Amphibian Oocytes, Mammalian Cell Cultures, Yeast, and Plants.
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两栖类卵母细胞、哺乳动物细胞培养物、酵母和植物的核孔复合物的扫描电子显微镜 (SEM) 和免疫 SEM。
DOI：
10.1007/978-1-0716-2337-4_27
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Goldberg MW
中科院分区：
文献类型：
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作者：
Goldberg MW
扫描电子显微镜（SEM）可以对许多生物和模型系统的核孔复合物（NPC）表面结构进行成像。使用场发射扫描电镜，这是一种中分辨率技术，可以直接成像各种组件的组织细节。有些成分，如NPC篮和细胞质细丝，很难用其他方式可视化。蛋白质成分可通过免疫金标记进行鉴定。任何可以暴露的表面都可以通过扫描电镜进行潜在的研究。本文给出了几种重叠的扫描电镜样品制备和免疫金标记NPCs的方案。详细介绍了样品制备、固定、免疫金标记、干燥、金属涂层和成像的各种参数，这些参数针对不同类型的标本和所需的终点进行了优化。
Scanning electron microscopy (SEM) can be used to image nuclear pore complex (NPC) surface structure of from a number of organisms and model systems. With a field emission SEM, this is a medium resolution technique where details of the organization of various components can be directly imaged. Some components, such as the NPC baskets and cytoplasmic filaments, are difficult to visualize in any other way. Protein components can be identified by immunogold labeling. Any surface that can be exposed can potentially be studied by SEM. Several overlapping protocols for SEM sample preparation and immunogold labeling of NPCs are given here. Various parameters for sample preparation, fixation, immunogold labeling, drying, metal coating, and imaging are detailed which have been optimized for different types of specimens and desired endpoints.