Exploring Membrane Lipid and Protein Diffusion by FRAP

  • Parijat Sarkar
  • Amitabha ChattopadhyayEmail author
Part of the Springer Protocols Handbooks book series (SPH)


Knowledge of membrane dynamics is crucial since it allows us to understand membrane function. Fluorescence recovery after photobleaching (FRAP) is a widely used technique to monitor diffusion of lipids and proteins in biological membranes. We outline here general aspects of FRAP, followed by a step-by-step guide to carry out FRAP measurements for exploring diffusion of fluorescently labeled lipids and proteins in membranes of attached cells and membranes of Candida albicans. In this process, we have provided detailed hands-on tips, judicious use of which would ensure reliability and quality of acquired FRAP data and associated analysis.


FRAP Confocal microscopy Lateral diffusion Mobile fraction Diffusion coefficient DiI GFP 



A.C. gratefully acknowledges support from SERB Distinguished Fellowship (Department of Science and Technology, Govt. of India). P.S. thanks the Council of Scientific and Industrial Research for the award of a Shyama Prasad Mukherjee Fellowship. A.C. is a Distinguished Visiting Professor at the Indian Institute of Technology Bombay (Mumbai), Adjunct Professor at the RMIT University (Melbourne, Australia), Tata Institute of Fundamental Research (Mumbai), and Indian Institute of Science Education and Research (Kolkata), and an Honorary Professor at the Jawaharlal Nehru Centre for Advanced Scientific Research (Bengaluru). We thank members of the Chattopadhyay laboratory for their comments and discussions.

Conflicts of Interest

There are no conflicts of interest to declare.


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.CSIR-Centre for Cellular and Molecular BiologyHyderabadIndia

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