Reflections on FRET imaging: formalism, probes, and implementation

E. JARES-ERIJMAN, THOMAS JOVIN

FRET and FLIM Imaging Techniques , Laboratory Techniques in Biochemistry and Molecular Biology

ACADEMIC PRESS

Lugar: Burlington; Año: 2009; p. 457 - 517

Imaging based on FRET can exploit all the modalities of fluorescence phenomena to provide information about the spatial distribution and states of association and function of biomolecules in the cellular con- text. In this chapter, we deal with fundamental aspects of the FRET formalism in the search for more direct and robust determinations under certain circumstances. A ‘‘FRET calculator’’ is introduced that permits athorough exploration of these principles even under conditions of extreme nonlinearity, that is, saturation of donor and/or acceptor excited states. The implementation of numerous new strategies for FRET determinations requires new ‘‘hardware,’’ which we briefly review, as well as ‘‘software,’’ both in the literal sense and in reference to new organic and nanoparticle probes. We supply a comprehensive tabulation of new and potential expression systems, most of which need to be explored systematically in and for FRET. A final attempt at ‘‘crystal‐balling,’’ the future prospects for FRET technology and its applications emphasizes the obvious: it is difficult to make predictions except for stating that images will get finer, faster, and more multidimensional. There are 149 citations from the literature.