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Measurement of Three-Dimensional Anisotropic Diffusion by Multiphoton Fluorescence Recovery after Photobleaching

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Abstract

The multiphoton fluorescence recovery after photobleaching (MP-FRAP) technique has been developed to measure the three-dimensional (3D) solute diffusion within biological systems. However, current 3D MP-FRAP models are based on isotropic diffusion and spatial domain analysis. The 3D anisotropic diffusion and frequency domain analysis for MP-FRAP measurements are rarely studied. In this study, a new technique is demonstrated for the quantitative and non-destructive determination of 3D anisotropic solute diffusion tensors within biological fibrosis tissues by multiphoton photobleaching and spatial Fourier analysis (SFA). Compared to the spatial domain analysis based MP-FRAP techniques, this SFA-based method has the capability for determining the 3D anisotropic diffusion tensors as well as the flexibility for satisfying initial and boundary conditions. First, a close-form solution of the 3D anisotropic diffusion equation is derived by solely using SFA. Next, this new method is validated by computer-simulated MP-FRAP experiments with pre-defined 3D anisotropic diffusion tensors as well as experimental diffusion measurements of FITC-Dextran (FD) molecules in aqueous glycerol solutions. Finally, this MP-FRAP technique is applied to the measurement of 3D anisotropic diffusion tensors of FD molecules within porcine tendon tissues. This study provides a new tool for complete determination of 3D anisotropic solute diffusion tensor in biological tissues.

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Acknowledgments

This project was supported by NIH Grants DE021134, DE018741, and AR055775, a NSF RII Grant predoctoral fellowship (EPS-0903795) to CS, and a NSF Graduate Research Fellowship to SEC.

Conflict of interest

None of the authors of this paper have a conflict of interest that might be construed as affecting the conduct or reporting of the work presented.

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Authors and Affiliations

  1. CU-MUSC Bioengineering Program, Department of Bioengineering, Clemson University, 173 Ashley Avenue, P.O. Box 250508, Charleston, SC, 29425, USA

    Changcheng Shi, Sarah E. Cisewski & Hai Yao

  2. Division of Nephrology, Department of Medicine, Medical University of South Carolina, Charleston, SC, USA

    P. Darwin Bell

  3. Department of Craniofacial Biology, Medical University of South Carolina, Charleston, SC, USA

    Hai Yao

Authors
  1. Changcheng Shi
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  2. Sarah E. Cisewski
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  3. P. Darwin Bell
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  4. Hai Yao
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Correspondence to Hai Yao.

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Associate Editor Sean S. Kohles oversaw the review of this article.

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Shi, C., Cisewski, S.E., Bell, P.D. et al. Measurement of Three-Dimensional Anisotropic Diffusion by Multiphoton Fluorescence Recovery after Photobleaching. Ann Biomed Eng 42, 555–565 (2014). https://doi.org/10.1007/s10439-013-0939-7

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  • DOI: https://doi.org/10.1007/s10439-013-0939-7

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