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Biophys. J. BioFAST: First Published May 13, 2005. doi:10.1529/biophysj.104.053652
© 2005 by the Biophysical Society.

A more recent version of this article appeared on August 1, 2005.
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SPECTROSCOPY, IMAGING, OTHER TECHNIQUES

Determining diffusion coefficients in inhomogeneous tissues using fluorescence recovery after photobleaching

Yvonne Hendrika Sniekers 1 and Corrinus Cornelis van Donkelaar 1*

1 Eindhoven University of Technology

* To whom correspondence should be addressed. E-mail: c.c.v.donkelaar{at}tue.nl.

Submitted on September 27, 2004
Revised on November 5, 2004
Accepted on 6 May 2005


  Abstract
Diffusion plays an important role in the transport of nutrients and signaling molecules. Diffusion coefficients (D) can be measured by fluorescence recovery after photo bleaching (FRAP). Available methods to analyze FRAP data, however, assume homogeneity in the environment of the bleached area and neglect geometrical restrictions to diffusion. Hence, diffusion coefficients in inhomogeneous materials, such as most biological tissues, cannot be assessed accurately. In this study, a new method for analyzing data from FRAP measurements has been developed, which is applicable to inhomogeneous tissues. It is based on a fitting procedure of the intensity recovery after photobleaching with a two-dimensional finite element analysis, which includes Fick's law for diffusion. The FE analysis can account for distinctive diffusivity in predefined zones, which allows determining diffusion coefficients in inhomogeneous samples. The method is validated theoretically and experimentally in both homogeneous and inhomogeneous tissues, subsequently applied to the proliferation zone of the growth plate. Finally, the importance of accounting for inhomogeneities, for appropriate assessment of diffusivity in inhomogeneous tissues, is illustrated.

Key Words: FRAP, cartilage, diffusivity, growth plate, signaling, transport




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Copyright © 2005 by the Biophysical Society.