Effect of fluorescence quenching by stimulated emission on the spectral properties of a solvent-sensitive fluorophore

Ignacy Gryczynski, Józef Kuśba, Zygmunt Gryczynski, Henryk Malak, Joseph R. Lakowicz

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We examined the emission spectra and wavelength-dependent anisotropies of the solvent-sensitive fluorophore 4-(dimethylamino)-4′-cyanostilbene (DCS) under condition of fluorescence quenching by stimulated emission. The sample was illuminated with a train of 10 ps pulses at 285 nm, and a train of stimulating pulses at 570 nm which were delayed by a time td relative to the excitation. Stimulated emission of DCS was demonstrated to occur by observation of gain in the long-wavelength beam. Illumination on the long-wavelength side of the emission spectrum with the long-wavelength time-delayed pulses resulted in a blue shift of the emission spectrum, and a progressive decrease of the emission anisotropy as the observation wavelength increased toward the stimulating wavelength. The spectral shifts and wavelength-dependent anisotropies of DCS were more pronounced in more viscous solvents where spectral relaxation is incomplete during the excited state lifetime. Light quenching of DCS in a low-viscosity solvent revealed no spectral shifts or wavelength-dependent anisotropies. Control measurements using acridine orange, which is relatively insensitive to solvent polarity, did not show any spectral shift or wavelength-dependent anisotropy with light quenching. The data for DCS can be explained by the presence of a time-dependent spectral shift and wavelength-selective quenching of the longer wavelength emission. In this model the relaxed state is formed following excitation of the unrelaxed state, and the relaxed state is preferentially quenched by long-wavelength illumination. Comparison of the data with model calculations indicates the presence of at least two spectral relaxation times. These results demonstrate that light quenching by stimulated emission acts selectively based on overlap of the stimulating wavelength with the emission spectrum. Observation of the emission spectrum in the presence of time-delayed and power-controlled long-wavelength pulses can be used to study time-dependent excited-state processes.

Original languageEnglish
Pages (from-to)10135-10144
Number of pages10
JournalJournal of Physical Chemistry
Issue number24
StatePublished - 13 Jun 1996


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