FRET efficiency is a measure of the energy transfer between the donor and acceptor fluorophores and is crucial for interpreting the results of a FRET experiment. It can be calculated using several methods:
Acceptor Photobleaching: This involves bleaching the acceptor fluorophore and observing the increase in donor fluorescence, which indicates the efficiency of energy transfer. Sensitized Emission: This method measures the increase in acceptor fluorescence due to energy transfer from the excited donor fluorophore. Fluorescence Lifetime Imaging Microscopy (FLIM): This technique measures the change in donor fluorescence lifetime, which decreases as FRET efficiency increases.
