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High-Resolution Imaging of Cellular Proteins pp 125–151Cite as

Using FRAP or FRAPA to Visualize the Movement of Fluorescently Labeled Proteins or Cellular Organelles in Live Cultured Neurons Transformed with Adeno-Associated Viruses

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1474))

Abstract

Fluorescence recovery after photobleaching (FRAP) and fluorescence redistribution after photoactivation (FRAPA) are complementary methods used to gauge the movement of proteins or sub-resolution organelles within cells. Using these methods we can determine the nature of the movement of labeled particles, whether it is random, constrained, or active, the coefficient of diffusion if applicable, binding and unbinding constants, and the direction of active transport. These two techniques have been extensively utilized to probe the cell biology of neurons. A practical outline of FRAP and FRAPA in cultured neurons is presented, including the preparation of the neurons and their infection with adeno-associated viral vectors. Considerations in planning such experiments are provided.

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Acknowledgement

This work was supported by Israel Science Foundation (ISF) grant 1427/12 (DG).

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Authors and Affiliations

  1. Department of Physiology and Cell Biology, Faculty of Health Sciences and Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, Beer-Sheva, 8410501, Israel

    Yaara Tevet & Daniel Gitler

Authors
  1. Yaara Tevet
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  2. Daniel Gitler
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Corresponding author

Correspondence to Daniel Gitler .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, The University of Memphis, Memphis, Tennessee, USA

    Steven D. Schwartzbach

  2. Department of Biological Sciences, The University of Memphis, Memphis, Tennessee, USA

    Omar Skalli

  3. Department of Anatomy, Universidad Central del Caribe, Bayamon, Puerto Rico, USA

    Thomas Schikorski

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Tevet, Y., Gitler, D. (2016). Using FRAP or FRAPA to Visualize the Movement of Fluorescently Labeled Proteins or Cellular Organelles in Live Cultured Neurons Transformed with Adeno-Associated Viruses. In: Schwartzbach, S., Skalli, O., Schikorski, T. (eds) High-Resolution Imaging of Cellular Proteins. Methods in Molecular Biology, vol 1474. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6352-2_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6352-2_8

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6350-8

  • Online ISBN: 978-1-4939-6352-2

  • eBook Packages: Springer Protocols

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