title: Electron tomography and immunogold labelling as tools to analyse de novo assembly of plant cell walls.
abstract: High-resolution imaging of the membranous intermediates and cytoskeletal arrays involved in the assembly of a new cell wall during plant cytokinesis requires state-of-the-art electron microscopy techniques. The combination of cryofixation/freeze-substitution methods with electron tomography (ET) has revealed amazing structural details of this unique cellular process. This chapter deals with the main steps associated with these imaging techniques: selection of samples suitable for studying plant cytokinesis, sample preparation by high-pressure freezing/freeze substitution, and ET of plastic sections. In addition, immunogold approaches for the identification of proteins and polysaccharides during cell wall assembly are discussed.
authors.affiliation: Department of Botany, University of Wisconsin, Madison, WI, USA. otegui@wisconsin.edu
authors.forename: Marisa S
authors.initials: MS
authors.lastname: Otegui
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure; D017360:Arabidopsis / Q000166:cytology* / Q000254:growth & development / Q000648:ultrastructure; D002473:Cell Wall / Q000378:metabolism*; D003198:Computer Simulation; D046508:Culture Techniques; D022162:Cytoplasmic Vesicles / Q000648:ultrastructure; D055032:Electron Microscope Tomography / Q000295:instrumentation / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008870:Microtubules / Q000648:ultrastructure; D053141:Organelle Size; D015388:Organelles / Q000648:ultrastructure; D016613:Plastic Embedding / Q000379:methods; D036226:Seedlings / Q000166:cytology* / Q000648:ultrastructure; D012984:Software