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Biomed Opt Express. 2012 Dec 1;3(12):3161-75. doi: 10.1364/BOE.3.003161. Epub 2012 Nov 8.

Quantitative tomographic imaging of intermolecular FRET in small animals.

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1
Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York. 12180, USA ; Currently with the Center for Molecular Imaging, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, Massachusetts 02215, USA.

Abstract

Forster resonance energy transfer (FRET) is a nonradiative transfer of energy between two fluorescent molecules (a donor and an acceptor) in nanometer range proximity. FRET imaging methods have been applied to proteomic studies and drug discovery applications based on intermolecular FRET efficiency measurements and stoichiometric measurements of FRET interaction as quantitative parameters of interest. Importantly, FRET provides information about biomolecular interactions at a molecular level, well beyond the diffraction limits of standard microscopy techniques. The application of FRET to small animal imaging will allow biomedical researchers to investigate physiological processes occurring at nanometer range in vivo as well as in situ. In this work a new method for the quantitative reconstruction of FRET measurements in small animals, incorporating a full-field tomographic acquisition system with a Monte Carlo based hierarchical reconstruction scheme, is described and validated in murine models. Our main objective is to estimate the relative concentration of two forms of donor species, i.e., a donor molecule involved in FRETing to an acceptor close by and a nonFRETing donor molecule.

KEYWORDS:

(110.6955) Tomographic imaging; (170.3650) Lifetime-based sensing; (170.6920) Time-resolved imaging

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