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Two photon-induced fluorescence intensity and anisotropy decays of diphenylhexatriene in solvents and lipid bilayers

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Abstract

We measured the fluorescence intensity and anisotropy decays of 1,6-diphenyl-1,3,5-hexatriene (DPH)-labeled membranes resulting from simultaneous two-photon excitation of fluorescence. Comparison of these two-photon data with the more usual one-photon measurements revealed that DPH displayed identical intensity decays, anisotropy decays, and order parameters for one- and two-photon excitation. While the anisotropy data are numerically distinct, they can be compared by use of the factor 10/7, which accounts for the two-photon versus one-photon photoselection. The increased time 0 anisotropy of DPH can result in increased resolution of complex anisotropy decays. Global analysis of the one- and two-photon data reveals consistency with a single apparent angle between the absorption and the emission oscillators. The global anisotropy analysis also suggests that, except for the photoselection factor, the anisotropy decays are the same for one-and two-photon excitation. This ideal behavior of DPH as a two-photon absorber, and its high two-photon cross section, makes DPH a potential probe for confocal two-photon microscopy and other systems where it is advantageous to use long-wavelength (680- to 760-nm) excitation.

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Authors and Affiliations

  1. Center for Fluorescence Spectroscopy, Department of Biological Chemistry, University of Maryland School of Medicine, 108 North Greene Streel, 21201, Baltimore, Maryland

    Joseph R. Lakowicz, Ignacy Gryczynski, Józef Kuśba & Eva Danielsen

  2. Department of Mathematics and Physics, Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871, Frederiksberg C, Denmark

    Eva Danielsen

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  1. Joseph R. Lakowicz
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  2. Ignacy Gryczynski
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  3. Józef Kuśba
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Lakowicz, J.R., Gryczynski, I., Kuśba, J. et al. Two photon-induced fluorescence intensity and anisotropy decays of diphenylhexatriene in solvents and lipid bilayers. J Fluoresc 2, 247–258 (1992). https://doi.org/10.1007/BF00865283

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  • DOI: https://doi.org/10.1007/BF00865283

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