Abstract
Bioluminescence imaging is a powerful technique that allows for deep-tissue analysis in living, intact organisms. However, in vivo optical imaging is compounded by difficulties due to light scattering and absorption. While light scattering is relatively difficult to overcome and compensate, light absorption by biological tissue is strongly dependent upon wavelength. For example, light absorption by mammalian tissue is highest in the blue-yellow part of the visible energy spectrum. Many natural bioluminescent molecules emit photonic energy in this range, thus in vivo optical detection of these molecules is primarily limited by absorption. This has driven efforts for probe development aimed to enhance photonic emission of red light that is absorbed much less by mammalian tissue using either direct genetic manipulation, and/or resonance energy transfer methods. Here we describe a recently identified alternative approach termed Fluorescence by Unbound Excitation from Luminescence (FUEL), where bioluminescent molecules are able to induce a fluorescent response from fluorescent nanoparticles through an epifluorescence mechanism, thereby significantly increasing both the total number of detectable photons as well as the number of red photons produced.
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Acknowledgements
Joe Dragavon is a Florence Gould Scholar of the Pasteur Foundation Postdoctoral Fellowship Program. The authors would like to extend their gratitude for financial support from the Pasteur Foundation of New York (to J.D., C.S.), the EU-FP7 Program “Automation” (to S.L.S.), the Institut Carnot Program 11 (to R.T., S.L.S.) and Project IMNOS (to R.T., S.L.S.), the Conny-Maeve Charitable Foundation (S.L.S.), the European Masters in Molecular Imaging (to I.T.), the Region Ile de France programs MODEXA (S.L.S.), SESAME (S.L.S.), and DimMalInf (S.L.S., R.T.), and the Institut Pasteur, Paris. Further, the authors would like to thank Bruno Baron of the Plate-Forme de Biophysique des Macromolécules et de leurs Interactions and Marie-Anne Nicola of the Plate-Forme d’Imagerie Dynamique for technical support and assistance, José Bengoechea and Herbert Schweizer for reagents, and Philippe Sansonetti for use of lab space, reagents, and equipment.
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Dragavon, J. et al. (2014). In Vitro and In Vivo Demonstrations of Fluorescence by Unbound Excitation from Luminescence (FUEL). In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-718-1_20
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DOI: https://doi.org/10.1007/978-1-62703-718-1_20
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