The polarization values (r) were calculated by the fluorescence data manager program using the Jablonski equation: where IVV and IVH are the vertical and horizontal fluorescence intensities, respectively to the vertical polarization of the excitation light beam. The factor G = IHV/IHH (grating correction factor) corrects the polarizing effects of the monochromator. The excitation wavelengths were 348 nm (DPH) and 340 nm (TMA-DPH) and the fluorescence emission was measured at 426 nm for DPH and 430 nm for TMA-DPH78 (link).
Based on the data obtained, the membrane ordering parameter was calculated using the equation47 : where r0 is the fluorescence anisotropy of DPH or TMA-DPH in the absence of any rotational motion of the probe. The theoretical value of r0 of DPH and TMA-DPH is 0.4.