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Fluorescence Microscopy of Microtubules in Cultured Cells

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Part of the Methods in Molecular Medicine™ book series (MIMM,volume 137)

Abstract

Cytoplasmic microtubules are noncovalent polymers of the protein tubulin. In the cells, the main function of microtubules is to provide tracks for organelle transport. Two experimental approaches based on fluorescence microscopy are commonly used to examine organization of microtubules in mammalian tissue culture cells. The first experimental approach involves indirect immunofluorescence staining of chemically fixed cells with tubulin antibody. Fluorescence microscopy of immunostained specimens allows the examination of the distribution of microtubules in the cytoplasm at the moment of fixation. The second experimental approach involves introduction of tubulin subunits covalently labeled with a fluorochrome into the cytoplasm of living cells. Time-lapse fluorescence microscopy of cells containing labeled tubulin subunits allows to examine changes in the spatial organization of microtubules in the cytoplasm and also to directly observe their behavior. In this chapter, we describe preparation of samples for fluorescence microscopy of microtubules.

Key Words

  • Microtubules
  • tubulin
  • fluorescence microscopy
  • microtubule dynamics
  • immunostaining
  • live cell imaging

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  • DOI: 10.1007/978-1-59745-442-1_6
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References

  1. Desai, A. and Mitchison, T. J. (1997) Microtubule polymerization dynamics. Ann. Rev. Cell Dev. Biol. 13, 83–117.

    CAS  CrossRef  Google Scholar 

  2. Komarova, J., Peloquin, J., and Borisy, G. (2004) Microinjection of fluorophore-labeled proteins, in Live Cell Imaging, (Goldman, R. D. and Spector, D. L., ed.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 67–86.

    Google Scholar 

  3. Goodson, H. V. and Wadsworth, P. (2004) Methods for expressing and analyzing GFP-tubulin and GFP-microtubule-associated proteins, in Live Cell Imaging, (Goldman, R. D. and Spector, D. L., ed.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 537–553.

    Google Scholar 

  4. Rusan, N. M., Fagerstrom, C. J., Yvon, A. M. C., and Wadsworth, P. (2001) Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-α tubulin. Mol. Biol. Cell 12, 971–980.

    CAS  PubMed  Google Scholar 

  5. Borisy, G. G., Marcum, J. M., Olmsted, J. B., Murphy, D. B., and Johnson, K. A. (1975) Purification of tubulin and associated high molecular weight proteins from porcine brain and characterization of microtubule assembly in vitro. Ann. N.Y. Acad. Sci. 253, 107–132.

    CAS  CrossRef  PubMed  Google Scholar 

  6. Rodionov, V. I., Nadezhdina, E. S., Peloquin, J., and Borisy, G. (2001) Digital fluorescence microscopy of cell cytoplasts with and without the centrosome. Methods Cell Biol. 67, 43–51.

    CAS  CrossRef  PubMed  Google Scholar 

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© 2007 Humana Press Inc.

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Semenova, I., Rodionov, V. (2007). Fluorescence Microscopy of Microtubules in Cultured Cells. In: Zhou, J. (eds) Microtubule Protocols. Methods in Molecular Medicine™, vol 137. Humana Press. https://doi.org/10.1007/978-1-59745-442-1_6

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  • DOI: https://doi.org/10.1007/978-1-59745-442-1_6

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-642-9

  • Online ISBN: 978-1-59745-442-1

  • eBook Packages: Springer Protocols