Cell Migration pp 387-402 | Cite as

Fluorescence Recovery After Photobleaching

  • Alex Carisey
  • Matthew Stroud
  • Ricky Tsang
  • Christoph BallestremEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 769)


This chapter describes the use of microscope-based fluorescence recovery after photobleaching (FRAP). To quantify the dynamics of proteins within a subcellular compartment, we first outline the general aspects of FRAP experiments and then provide a detailed protocol of how to measure and analyse the most important parameters of FRAP experiments such as mobile fraction and half-time of recovery.

Key words

Fluorescence recovery after photobleaching Fluorescence microscopy Green fluorescent protein Half-time of recovery Mobile fraction Diffusion Binding reaction kinetics Focal adhesions Vinculin 



We would like to thank Dr. Janet Askari for critical reading of the manuscript. CB acknowledges BBSRC (BB/G004552/1). The Bioimaging Facility microscopes used in this study were purchased with grants from BBSRC, Wellcome Trust and the University of Manchester Strategic Fund.


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Alex Carisey
  • Matthew Stroud
  • Ricky Tsang
  • Christoph Ballestrem
    • 1
    Email author
  1. 1.Faculty of Life SciencesThe University of ManchesterManchesterUK

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