Polishchuk R S, Mironov A A
Consorzio Mario Negri Sud, S. Maria Imbara (Chieti), Italy.
Curr Protoc Cell Biol. 2001 Aug;Chapter 4:Unit 4.8. doi: 10.1002/0471143030.cb0408s11.
This unit describes newly developed methods that allow the examination of living cells by time-lapse analysis with the subsequent identification of the just-observed organelle under an electron microscope. To understand how such cellular functions, such as intracellular traffic, cytokinesis, and cell migration, are organized and executed in vivo, it is most useful to observe living cells in real time with the spatial resolution afforded by electron microscopy (EM). Most suitable for this is a conceptually simple, yet powerful, method called correlative video light/electron microscopy (CVLEM), by which observations of the in vivo dynamics and the ultrastructure of intracellular objects can indeed be combined to achieve the above-mentioned result. This unit describes this methodology, illustrates the type of questions that the CVLEM approach was designed to address, and discusses the expertise required for successful application of the technique.
本单元介绍了新开发的方法，这些方法允许通过延时分析来检查活细胞，并随后在电子显微镜下识别刚刚观察到的细胞器。为了理解诸如细胞内运输、胞质分裂和细胞迁移等细胞功能在体内是如何组织和执行的，利用电子显微镜（EM）提供的空间分辨率实时观察活细胞是非常有用的。最适合于此的是一种概念上简单但功能强大的方法，称为相关视频光/电子显微镜（CVLEM），通过这种方法，可以将对细胞内物体的体内动力学和超微结构的观察结合起来，以实现上述结果。本单元描述了这种方法，说明了CVLEM方法旨在解决的问题类型，并讨论了成功应用该技术所需的专业知识。
