A new double-exponential FLIM-FRET approach obtains FRET parameters from a single lifetime image of the donor.
By using the lifetime of the non-interacting donor fraction as a reference the method avoids the need for reference data from separate donor-only cells. This not only makes the technique easy to use but also prevents differences in the metabolic state of the cells from influencing the FRET results. The technique simultaneously delivers the classic FRET efficiency, the pure FRET efficiency of the interacting donor, and the relative amount of interacting donor. Please see Application Note for details.
