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Two-Photon Activated Two-Photon Fluorescence and Binding of Azidocoumarin in a Gelatin Matrix

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Abstract

We study the creation of fluorescence patterns inside a gelatin gel by way of two-photon photoactivation of 7-azido-4-trifluoromethyl-1,2-benzopyrone (azidocomarin 151) contained in the gel matrix. As ultrafast light pulses are focused into the gel, onset of two-photon fluorescence, highly nonlinear in the applied optical power, is observed as azidocoumarin is converted into a fluorescent dye that binds to the gelatin. We fit the time dependence of the fluorescence to a model that incorporates the competition between coumarin photoactivation and photobleaching as well as the gradual degradation of the gel when it is exposed to the high intensity laser light. The model predicts that the initial rate of fluorescence onset should scale as the P 4, where P is laser power, while the signal at long exposure time should scale as P 3/2. The observed exponents are 4.18 and 1.34, respectively. The model allows us to estimate the cross section and quantum yield of two-photon induced photobleaching of azidocoumarin 151. The numerous technical uses of gelatin and the collagen from which it derives in areas ranging from photography to tissue engineering provide possible applications for the techniques described in this paper.

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Acknowledgements

This work was funded by a grant from the National Science Foundation (agreement CBET-0756693), the Institute for Critical Technology and Applied Science (ICTAS) and by a grant from the Thomas F. Jeffress and Kate M. Jeffress Memorial Trust.

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  1. Department of Physics, Virginia Tech, 910 Drillfield Drive, Blacksburg, VA, 24061, USA

    Stefan V. Stoianov & Hans D. Robinson

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  1. Stefan V. Stoianov
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  2. Hans D. Robinson
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Correspondence to Hans D. Robinson.

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Stoianov, S.V., Robinson, H.D. Two-Photon Activated Two-Photon Fluorescence and Binding of Azidocoumarin in a Gelatin Matrix. J Fluoresc 22, 1291–1300 (2012). https://doi.org/10.1007/s10895-012-1071-z

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