| Abstract: |
The phenomenon of fluorescence resonance energy transfer (FRET) was first described by Theodor Forster in 1948. In this chapter, the authors introduce the reader to the photophysics underlying FRET and present a brief general overview of the types of biological measurements enabled by the technique. FRET efficiency is inversely proportional to the sixth power of the distance between a donor and acceptor, and therefore is extremely sensitive to their intermolecular separation. A wide array of phenomena can be measured using FRET, and they tend to fall into either of two main categories: homogeneous in vitro assays performed in a spectrofluorimeter or microplate reader and live-cell FRET imaging performed by fluorescence microscopy. FRET measurements are not limited to in vitro assays. Cell-permeable FRET probes as well as the green, cyan, and yellow fluorescent proteins (GFP, CFP, and YFP), introduced in cells by DNA transfection, have permitted the study of dynamic events involving changes in the proximity of macromolecules in live cells. © 2022 Taylor & Francis Group, LLC. |
