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Immunogold Labeling and Electron Tomography of Plant Endosomes

  • Alexandra Chanoca
  • Marisa S. OteguiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1209)

Abstract

High-resolution imaging of endosomal compartments and associated organelles can be achieved using state-of-the-art electron microscopy techniques, such as the combination of cryofixation/freeze-substitution for sample processing and electron tomography for three-dimensional (3D) analysis. This chapter deals with the main steps associated with these imaging techniques: selection of samples suitable for studying plant endosomes, sample preparation by high-pressure freezing/freeze-substitution, and electron tomography of plastic sections. In addition, immunogold approaches for identification of subcellular localization of endosomal and cargo proteins are also discussed.

Key words

Electron tomography Endosomes Trans-Golgi network Multivesicular body Cryofixation Immunolabeling 

Notes

Acknowledgements

This work was supported by NSF grant MCB1157824 to M.S.O.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of BotanyUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of GeneticsUniversity of Wisconsin-MadisonMadisonUSA

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