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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 4, Iss. 11 — Oct. 21, 2009

Three-dimensional imaging of Förster resonance energy transfer in heterogeneous turbid media by tomographic fluorescent lifetime imaging

James McGinty, Vadim Y. Soloviev, Khadija B. Tahir, Romain Laine, Daniel W. Stuckey, Joseph V. Hajnal, Alessandro Sardini, Paul M.W. French, and Simon R. Arridge  »View Author Affiliations


Optics Letters, Vol. 34, Issue 18, pp. 2772-2774 (2009)
http://dx.doi.org/10.1364/OL.34.002772


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Abstract

We report a three-dimensional time-resolved tomographic imaging technique for localizing protein–protein interaction and protein conformational changes in turbid media based on Förster resonant energy-transfer read out using fluorescence lifetime. This application of “tomoFRET” employs an inverse scattering algorithm utilizing the diffusion approximation to the radiative-transfer equation applied to a large tomographic data set of time-gated images. The approach is demonstrated by imaging a highly scattering cylindrical phantom within which are two thin wells containing cytosol preparations of HEK293 cells expressing TN-L15, a cytosolic genetically encoded calcium Förster resonant energy-transfer sensor. A 10 mM calcium chloride solution was added to one of the wells, inducing a protein conformation change upon binding to TN-L15, resulting in Förster resonant energy transfer and a corresponding decrease in the donor fluorescence lifetime. We successfully reconstruct spatially resolved maps of the resulting fluorescence lifetime distribution as well as of the quantum efficiency, absorption, and scattering coefficients.

© 2009 Optical Society of America

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.3010) Medical optics and biotechnology : Image reconstruction techniques
(290.0290) Scattering : Scattering
(290.7050) Scattering : Turbid media

ToC Category:
Scattering

History
Original Manuscript: May 28, 2009
Revised Manuscript: July 26, 2009
Manuscript Accepted: August 8, 2009
Published: September 9, 2009

Virtual Issues
Vol. 4, Iss. 11 Virtual Journal for Biomedical Optics

Citation
James McGinty, Vadim Y. Soloviev, Khadija B. Tahir, Romain Laine, Daniel W. Stuckey, Joseph V. Hajnal, Alessandro Sardini, Paul M. W. French, and Simon R. Arridge, "Three-dimensional imaging of Förster resonance energy transfer in heterogeneous turbid media by tomographic fluorescent lifetime imaging," Opt. Lett. 34, 2772-2774 (2009)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=ol-34-18-2772

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