Abstract
Upon excitation with polarized light the emission from fluorescent samples is also polarized. This polarization is a result of the photoselection of fluorophores according to their orientation relative to the direction of the polarized excitation. The emission can be depolarized by a number of phenomena, the relative importance of which depends upon the sample under investigation. Rotational diffusion of fluorophores is one common cause of depolarization. The polarization or anisotropy measurements reveal the average angular displacement of the fluorophore which occurs between absorption and subsequent emission of a photon. This angular displacement is dependent upon the rate and extent of rotational diffusion during the lifetime of the excited state. These diffusive motions in turn depend upon the viscosity of the solvent and the size and shape of the diffusing species. For example, for a fluorophore dissolved in a solvent, the rotational rate of the fluorophore is dependent upon the viscous drag imposed on the fluorophore by the solvent. As a result, a change in solvent viscosity will result in a change in fluorescence anisotropy.
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© 1983 Plenum Press, New York
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Lakowicz, J.R. (1983). Fluorescence Polarization. In: Principles of Fluorescence Spectroscopy. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-7658-7_5
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DOI: https://doi.org/10.1007/978-1-4615-7658-7_5
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