Scanning electron microscopy (SEM) takes advantage of distinct detectors to visualise secondary and back-scattering electrons. Here, we report an integrated approach that relies on these two detection methods to simultaneously acquire correlated information on plasma membrane topography and curvature-sensitive cytosolic protein localization in intact cell samples. We further provide detailed preparation and staining protocols, as well as a thorough example-based discussion for imaging optimisation. Collectively, the presented method enables rapid and precise analysis of cytosolic proteins adjacent to cellular membranes with a resolution of ~100 nm, without time-consuming preparations or errors induced by sequential visualisation present in fluorescence-based correlative approaches.

中文翻译：

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通过相关免疫金相SEM并行获取培养细胞中的质膜超微结构和胞浆蛋白定位。

扫描电子显微镜（SEM）利用独特的检测器来可视化二次电子和反向散射电子。在这里，我们报告了一种综合的方法，该方法依赖于这两种检测方法来同时获取完整细胞样品中质膜形貌和曲率敏感性细胞溶质蛋白定位的相关信息。我们还提供详细的准备和染色方案，以及基于实例的彻底讨论，以优化成像。总体而言，本文提出的方法能够快速，精确地分析细胞膜附近的胞质蛋白，分辨率约为100 nm，而无需耗时的准备工作或因基于荧光的相关方法中的顺序可视化而引起的错误。