Novel Video-FRAP method for the determination of anisotropic diffusion: Principles and optimization

Research output: Chapter in Book/Report/Conference proceedingConference contribution


Fluorescence recovery after photobleaching (FRAP) is an established technique for determining solute diffusivity in biological tissues. Recently, we proposed a novel fluorescence photobleaching method, based on two independent analyses of the video-FRAP image series: the fast Fourier transform (FFT) and the Karhunen-Loève transform (KLT). In this study, the principles of this method are illustrated, and its optimization is discussed. Computer simulated FRAP experiments were used for the optimization of the choice of frequencies used in FFT analysis. The sensitivity of the technique to image noise contamination was evaluated. Additionally, the method was experimentally validated by characterizing fluorescein diffusivity in human annulus fibrosus. The results were compared to diffusivity data obtained by other methods. This study provides new insights into the optimization of data analysis of video-FRAP experiments. Furthermore, the experimental results verified the applicability of the method for the determination of solute diffusivity in biological media.

Original languageEnglish (US)
Title of host publication25th Southern Biomedical Engineering Conference 2009
Number of pages2
StatePublished - Nov 6 2009
Event25th Southern Biomedical Engineering Conference 2009 - Miami, FL, United States
Duration: May 15 2009May 17 2009

Publication series

NameIFMBE Proceedings
ISSN (Print)1680-0737


Other25th Southern Biomedical Engineering Conference 2009
CountryUnited States
CityMiami, FL



  • Annulus Fibrosus (AF)
  • Fast Fourier Transform (FFT)
  • Gaussian noise

ASJC Scopus subject areas

  • Biomedical Engineering
  • Bioengineering

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