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Strategies for Imaging Microtubules in Plant Cells

  • Andreas Holzinger
  • Eiko Kawamura
  • Geoffrey O. Wasteneys
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 586)

Summary

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.

Key words

Alexa Microtubules Dynein EB1 GFP, Kinesin MAP4 MBD mor1-1 

Notes

Acknowledgments

This chapter has been supported in part by a grant from the Universitätszentrum Obergurgl, University of Innsbruck, Austria to AH and funding from the Natural Sciences and Engineering Research Council and the Canadian Institutes of Health Research to GOW. The Thieme Verlag KG is acknowledged for their kind permission to reproduce Fig. 1b, c and Elsevier for reproduction of Fig. 7.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Andreas Holzinger
    • 1
  • Eiko Kawamura
    • 2
  • Geoffrey O. Wasteneys
    • 2
  1. 1.Institute of Botany, Department of Physiology and Cell PhysiologyUniversity of InnsbruckInnsbruckAustria
  2. 2.Department of BiologyUniversity of British ColumbiaVancouverCanada

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