Correlative light and electron microscopy (CLEM) methods combined with live imaging can be applied to understand the dynamics of organelles. Although recent advances in cell biology and light microscopy have helped visualize the details of organelle activities, observing their ultrastructure or organization of surrounding microenvironments is a challenging task. Therefore, CLEM, which allows us to observe the same area as an optical microscope with an electron microscope, has become a key technique in cell biology. Unfortunately, most CLEM methods have technical drawbacks, and many researchers face difficulties in applying CLEM methods. Here, we propose a live-3-dimensional (3D) CLEM method, combined with a 3D reconstruction technique using FIB-SEM tomography, as a solution to such technical barriers. We review our method, the associated technical limitations, and the options considered to perform live CLEM.

中文翻译：

---

使用 FIB-SEM 的光显微实时成像和电子显微 3D 结构之间细胞器动力学的相关性

相关光和电子显微镜 (CLEM) 方法与实时成像相结合，可用于了解细胞器的动态。尽管细胞生物学和光学显微镜的最新进展有助于可视化细胞器活动的细节，但观察它们的超微结构或周围微环境的组织是一项具有挑战性的任务。因此，使我们能够用电子显微镜观察与光学显微镜相同的区域的 CLEM 已成为细胞生物学中的一项关键技术。不幸的是，大多数 CLEM 方法都存在技术缺陷，许多研究人员在应用 CLEM 方法时面临困难。在这里，我们提出了一种实时 3 维 (3D) CLEM 方法，结合使用 FIB-SEM 断层扫描的 3D 重建技术，作为解决此类技术障碍的方法。我们回顾我们的方法，