| Literature DB >> 26498784 |
Katherine Celler1, Miki Fujita1, Eiko Kawamura2, Chris Ambrose3, Klaus Herburger4, Andreas Holzinger5, Geoffrey O Wasteneys1.
Abstract
Microtubules (MTs) are required throughout plant development for a wide variety of processes, and different strategies have evolved to visualize and analyze them. This chapter provides specific methods that can be used to analyze microtubule organization and dynamic properties in plant systems and summarizes the advantages and limitations for each technique. We outline basic methods for preparing samples for immunofluorescence labeling, including an enzyme-based permeabilization method, and a freeze-shattering method, which generates microfractures in the cell wall to provide antibodies access to cells in cuticle-laden aerial organs such as leaves. We discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide chemical fixation, high-pressure freezing/freeze substitution, and post-fixation staining protocols for preserving MTs for transmission electron microscopy and tomography.Entities:
Keywords: ARK1; Correlative light and electron microscopy; EB1; Electron tomography; GFP; Immunofluorescence; Kinesin; Live cell imaging; MAP4; MBD; MOR1; Microtubules
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Year: 2016 PMID： 26498784 PMCID： PMC4874466 DOI： 10.1007/978-1-4939-3124-8_8
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745