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Immunogold Labeling and Electron Tomography of Plant Endosomes

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Plant Endosomes

Part of the book series: Methods in Molecular Biology ((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.

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Acknowledgements

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

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Authors and Affiliations

  1. Department of Botany, University of Wisconsin-Madison, B119 Birge Hall, 430 Lincoln Drive, Madison, WI, 53706, USA

    Alexandra Chanoca & Marisa S. Otegui

  2. Department of Genetics, University of Wisconsin-Madison, 425-g Henry Mall, Madison, WI, 53706, USA

    Marisa S. Otegui

Authors
  1. Alexandra Chanoca
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  2. Marisa S. Otegui
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Corresponding author

Correspondence to Marisa S. Otegui .

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Editors and Affiliations

  1. University of Wisconsin-Madison, Madison, Wisconsin, USA

    Marisa S. Otegui

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© 2014 Springer Science+Business Media New York

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Chanoca, A., Otegui, M.S. (2014). Immunogold Labeling and Electron Tomography of Plant Endosomes. In: Otegui, M. (eds) Plant Endosomes. Methods in Molecular Biology, vol 1209. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1420-3_5

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  • DOI: https://doi.org/10.1007/978-1-4939-1420-3_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1419-7

  • Online ISBN: 978-1-4939-1420-3

  • eBook Packages: Springer Protocols

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