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利用相关光电子显微镜分析高尔基体复合物的功能。

Analysis of Golgi complex function using correlative light-electron microscopy.

作者信息

Polishchuk Elena V, Polishchuk Roman S

机构信息

Telethon Institute of Genetics and Medicine, Naples, Italy.

出版信息

Methods Cell Biol. 2013;118:243-58. doi: 10.1016/B978-0-12-417164-0.00015-X.

Abstract

Though it has been studied for more than 100 years, the Golgi complex remains a fascinating organelle for cell biologists. This is because it is a unique intracellular structure with a list of well-known functions that continues to grow with the discovery of new genes and the development of novel advanced techniques. The Golgi apparatus has been carefully characterized in different organisms, tissues, and cell types both by light and by electron microscopy. However, the new quality step in Golgi research began with the development of correlative light-electron microscopy (CLEM). This innovative method allows visualizing the fate of the same organelle of interest at any moment of its life span first by video imaging in live cells and then by immunoelectron microscopy and 3D reconstruction. Here, we will present the historical overview of different Golgi-associated processes characterized by CLEM and describe the detailed protocol of this complex yet valuable technique.

摘要

尽管高尔基体已经被研究了100多年,但它仍然是细胞生物学家感兴趣的细胞器。这是因为它是一种独特的细胞内结构,具有一系列众所周知的功能,并且随着新基因的发现和新型先进技术的发展,其功能列表还在不断增加。通过光学显微镜和电子显微镜,高尔基体已经在不同的生物体、组织和细胞类型中得到了详细的表征。然而,高尔基体研究的新突破始于相关光电子显微镜(CLEM)的发展。这种创新方法能够在感兴趣的细胞器生命周期的任何时刻,首先通过活细胞视频成像,然后通过免疫电子显微镜和三维重建来观察其命运。在这里,我们将介绍通过CLEM表征的不同高尔基体相关过程的历史概况,并描述这种复杂但有价值的技术的详细方案。

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