Analysis of Golgi complex function using correlative light-electron microscopy

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.

Keywords: Correlative light-electron microscopy; Gold labeling; Golgi complex; Green fluorescent protein; Immunoelectron microscopy; Live-cell imaging; Preembedding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Dogs
  • Gold / chemistry
  • Golgi Apparatus / physiology*
  • Golgi Apparatus / ultrastructure*
  • Green Fluorescent Proteins / biosynthesis
  • Imaging, Three-Dimensional
  • Immunohistochemistry
  • Madin Darby Canine Kidney Cells
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Microscopy, Immunoelectron
  • Microtomy
  • Plastic Embedding
  • Staining and Labeling

Substances

  • Green Fluorescent Proteins
  • Gold