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Vol. 35 No. 3
May-June 2013

Making an imPACt | Recent IUPAC technical reports and recommendations that affect the many fields of pure and applied chemistry.
See also www.iupac.org/publications/pac

Fluorescence Anisotropy Measurements in Solution: Methods and Reference Materials (IUPAC Technical Report)

Marcel Ameloot, et al., Pure and Applied Chemistry, 2013
Vol. 85, No. 3, pp. 589–608

The measurement of stationary and time-resolved linearly (or plane) polarized fluorescence in solution is an important method of investigating the physical and chemical molecular properties of the electronic structure and solvent–solute interactions of small molecules, the conformation and dynamics of natural and synthetic macromolecules, and more. In addition, a growing number of very sensitive and specific analytical methods are based on the determination of changes in fluorescence polarization. The theoretical aspects underlying the emission of polarized fluorescence are presented with great detail in a number of general and specific textbooks and journal publications. Conversely, the practical aspects of the corresponding experimental techniques, including the description of the most frequent sources of systematic errors that corrupt the measurement of polarized fluorescence intensity, are quite dispersed in the literature. As a consequence, the authors of this paper present a comprehensive discussion of spectroscopic methods used currently in the measurement of fluorescence polarization, both steady-state and time-resolved, focusing on those practices that provide accurate and reproducible values of fluorescence polarization from liquid solutions. Methods that are specific for fluorescence microscopy and multiphoton applications will only be touched on briefly, as far as they have consequences for the availability and applicability of polarization standards.

http://dx.doi.org/10.1351/PAC-REP-11-11-12



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