From a stock of purified liposomes, 60 μl was pipetted into a disposable cuvette and mixed with 1140 µl of PBS. Then the measurement (in triplicates for each sample) of CF permeation through the membrane was carried out in a fluorescence spectrophotometer (Cary Eclipse, Agilent) in which the sample was heated to the desired temperature (30 °C, 40 °C and 50 °C), which was kept constant throughout the measurement. The following settings were used: excitation wavelength 490 nm, emission wavelength 522 nm, excitation slit: 2.5 and 2. of the fluorescence intensity at constant temperatures was measured. At the end of the experiment, 5 µl of ten times diluted TRITON X-100 ® was added to cause total micellization of the system, thus releasing all previously unreleased CF. The mechanism of this micellization is shown in Fig. and is based on molecular dynamics study . The measured fluorescence intensity dependence on time was then converted to CF concentration using a calibration curve.