Strategies for imaging microtubules in plant cells.
Microtubules are required throughout plant development for a wide variety of processes, and different strategies have been evolved to visualize them. This chapter summarizes the most effective of these methods and points out potential problems and pitfalls. We outline the freeze-shattering method for immunolabeling microtubules in aerial organs such as leaves that require mechanical permeabilization, discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide different fixation protocols for preserving MTs for transmission electron microscopy including chemical fixation, high pressure freezing/freeze substitution, and post-fixation staining procedures for transmission electron microscopy.
Holzinger
Andreas
A
Institute of Botany, Department of Physiology and Cell Physiology, University of Innsbruck, Innsbruck, Austria.
Kawamura
Eiko
E
Wasteneys
Geoffrey O
GO
D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000378:metabolism; D015925:Cryopreservation; D056890:Eukaryota / Q000166:cytology / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D005404:Fixatives; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure*; D059828:Plant Cells; D018515:Plant Leaves / Q000648:ultrastructure; D010944:Plants / Q000378:metabolism / Q000648:ultrastructure*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016610:Tissue Embedding; D016707:Tissue Fixation / Q000379:methods; D014404:Tubulin / Q000378:metabolism
D029681:Arabidopsis Proteins; D005404:Fixatives; C429183:MOR1 protein, Arabidopsis; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D014404:Tubulin; D049452:Green Fluorescent Proteins