Figure 4: Increase of fluorescence upon dequenching after dilution of the dyes.
From: Actual fusion efficiency in the lipid mixing assay - Comparison between nanodiscs and liposomes
Dequenching obtained from Eq. (2) using solutions containing liposomes with 1.5% (no dilution, d = 1), 0.75% (diluted twice, d = 2) and 0.19% (diluted 8 times, d = 8) NBD-DOPE and Rho-DOPE lipids. Dilution experiments were performed at 32 °C, 37 °C and 42 °C. The results do not depend on the temperature and are consistent with previous reports at 37 °C12 (crosses). The continuous line, represented between d = 1 and d = 100 corresponds to the fit obtained by using Eq. (3) between d = 1 and d = 8. Representing it until d = 100 was only done for the sake of clarity, to check that, indeed, this fit allowed to find that, from one point, further dilution does not lead to further dequenching, since the dyes are already sufficiently far from each other for FRET not to occur.
