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  • 1
    ISSN: 1573-4994
    Keywords: Fluorescence anisotropy decays ; frequency-domain fluorescence spectroscopy ; review
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract This didactic paper summarizes the mathematical expressions needed for analysis of fluorescence anisotropy decays from polarized frequency-domain fluorescence data. The observed values are the phase angle difference between the polarized components of the emission and the modulated anisotropy, which is the ratio of the polarized and amplitude-modulated components of the emission. This procedure requires a separate measurement of the intensity decay of the total emission. The expressions are suitable for any number of exponential components in both the intensity decay and the anisotropy decay. The formalism is generalized for global analysis of anisotropy decays measured at different excitation wavelengths and for different intensity decay times as the result of quenching. Additionally, we describe the expressions required for associated anisotropy decays, that is, anisotropy decays where each correlation time is associated with a decay time present in the anisotropy decay. And finally, we present expressions appropriate for distributions of correlation times. This article should serve as a reference for researchers using frequency-domain fluorometry.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-4994
    Keywords: Anisotropy ; 4-dimethylamino-4′-cyanostilbene ; light quenching ; time-resolved fluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract We examined the emission wavelength-dependent anisotropies of the solvent-sensitive fluorophore 4-dimethylamino-4′-cyanostilbene (DCS) under conditions of light quenching by polarized time-delayed quenching pulses. Illumination on the long-wavelength side of the emission spectrum with time-delayed light pulses resulted in a progressive decrease in the emission anisotropy as the observation wavelength increased toward the stimulating wavelength. The anisotropy changes of DCS were most wavelength dependent when spectral relaxation occurred during the excited-state lifetime. Light quenching of DCS in a low-viscosity solvent revealed no wavelength-dependent anisotropies. Control measurements using a solvent-insensitive fluorophore did not show any wavelength-dependent anisotropy with light quenching. The data for DCS can be explained by a model which allows wavelength-selective quenching of the long-wavelength emission formed by time-dependent spectral relaxation. These results indicate that polarized light quenching can be used to study systems which display multiple emissions and/or time-dependent spectral shifts.
    Type of Medium: Electronic Resource
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