Impact of emission anisotropy on fluorescence spectroscopy and FRET distance measurements.

ABSTRACT: The objective of this report is to provide a practical and improved method for estimating Förster resonance energy transfer distance measurement error due to unknown angles in the dipole orientation factor based on emission anisotropy measurements. We improve on the method of Dale et al. (1979), which has minor mistakes and is frequently interpreted in overly optimistic ways in the literature. To facilitate proper fluorescence intensity measurements, we also evaluated instrument parameters that could impact the measurement. The apparent fluorescence intensity of isotropic samples depends on the sample emission anisotropy, fluorometer geometry, and optical apertures. We separate parameters of the sample, and those of the cylindrically symmetric illumination source and detector in the equati

SUBMITTER: Ivanov V 

PROVIDER: S-EPMC2718156 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature