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Fluorescence Proteins and Time-Lapse Imaging of the Cytoskeleton

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The Cytoskeleton

Part of the book series: Neuromethods ((NM,volume 79))

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Abstract

Visualization of cytoskeletal dynamics in real time is of paramount interest in cell biological research. With the aid of fluorescent-cytoskeletal fusion proteins and enhancement of confocal laser scanning microscopes with high-end objectives and cell-incubation chambers, high-resolution time-lapse imaging is nowadays possible for long time periods. However, most of the cytoskeletal dynamics can be detected during short observation periods.

In this chapter, we provide a detailed description for time-lapse imaging of microtubules, neurofilaments, and microfilaments within primary neurons. We use two primary neuronal cell culture systems for the analysis of different aspects of cytoskeletal motion and organization: (1) dissociated dorsal root ganglia, which are highly practical to study cytoskeletal dynamics along their neurites or within the growth cone, and (2) cerebellar slice cultures, which are characterized by their organotypic morphology even after 30 days in vitro. In particular in these slice cultures Purkinje cells exhibit highly dynamic dendritic spines within a functional neuronal network.

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Authors and Affiliations

  1. Department of Cytology, Ruhr University of Bochum, Bochum, Germany

    Carsten Theiss & Karl Meller

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  1. Carsten Theiss
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  2. Karl Meller
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Correspondence to Carsten Theiss .

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  1. Faculty of Medicine, Neuroanatomy & Molecular Brain Research, Ruhr-Universität Bochum, Universitätsstrasse 150, Bochum, 44801, Germany

    Rolf Dermietzel

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Theiss, C., Meller, K. (2013). Fluorescence Proteins and Time-Lapse Imaging of the Cytoskeleton. In: Dermietzel, R. (eds) The Cytoskeleton. Neuromethods, vol 79. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-266-7_1

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  • DOI: https://doi.org/10.1007/978-1-62703-266-7_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-265-0

  • Online ISBN: 978-1-62703-266-7

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