Fluorescence spectroscopy and energy transfer processes in biological systems

Abstract

This article is divided into three parts. In the first part we review the fundamental principles of fluorescence spectroscopy, starting with the consideration of fluorophores and of the characteristics of fluorescence spectroscopy. The processes of fluorescence quenching, fluorescence anisotropy, and resonance energy transfer are presented, together with the information they can provide. The techniques that produce absorption spectra, excitation spectra, fluorescence under continuous excitation and response to pulsed excitation are also examined. In the second part the basic interactions between atoms are introduced by considering first the static and then the dynamic effects of these interactions in a two-atom system ad in a linear chain of atoms. Subsequently the different types of interactions (multipolar electric and magnetic, and exchange) are examined. After a review of the different modes of excitation of a system containing both donors and acceptors, a statistical treatment of energy transfer is presented by considering first the case of energy transfer without migration among donors, and then the case when such migration occurs. In the third part the concepts presented in the second part are applied to distance distribution analysis and FRET (Fluorescence Resonance Energy Transfer) in biological systems. © 2011 Springer Science+Business Media B.V.