MEASURING FORSTER RESONANCE ENERGY TRANSFER WITH POLARIZED AND DEPOLARIZED LIGHT

摘要

The present application provides improved methods for assessing Förster resonance energy transfer (FRET) using polarized light. Specifically, the methods rely on measuring depolarized light emitted by fluorescent acceptor molecules. Depolarization of fluorescence emission is observed only in the presence of energy transfer. A simple detection strategy is described for fluorescence microscopy using laser scanning and wide field techniques. FRET between linked and unlinked green fluorescent proteins (GFPs) in living cells can be monitored with a larger dynamic range than in other techniques.