title: Immunogold labeling and electron tomography of plant endosomes.
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.
authors.affiliation: Department of Botany, University of Wisconsin-Madison, B119 Birge Hall, 430 Lincoln Drive, Madison, WI, 53706, USA.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Chanoca
authors.forename: Marisa S
authors.initials: MS
authors.lastname: Otegui
mesh_terms: D017360:Arabidopsis / Q000648:ultrastructure; D015925:Cryopreservation / Q000379:methods; D011992:Endosomes / Q000648:ultrastructure*; D008854:Microscopy, Electron / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D012639:Seeds / Q000648:ultrastructure; D021601:trans-Golgi Network / Q000648:ultrastructure*