Exploring the effects of sterols in model lipid membranes using single-molecule orientations.

Single-molecule orientations of the fluorescent lipid analogue BODIPY-PC doped into lipid monolayers and bilayers of DPPC are used to characterize the structure present in the films as a function of sterol content. Out-of-focus polarized total internal reflection fluorescence microscopy (PTIRF-M) measurements are used to characterize the single-molecule tilt angles with respect to the membrane normal. Tilt angle histograms for Langmuir-Blodgett monolayers of DPPC reveal bimodal distributions at all surface pressures studied. A linear dependence in the dye population oriented normal to the membrane plane with surface pressure is found and used to characterize the equivalent surface pressure of supported bilayers formed through vesicle fusion. These measurements reveal an equivalent surface pressure of approximately 23 mN/m, which is somewhat lower than the currently accepted value of approximately 30-35 mN/m.(1-7) The effect of cholesterol, ergosterol, and lanosterol on membrane structure is also compared between DPPC bilayers and monolayers transferred at approximately 23 mN/m. The addition of cholesterol leads to dramatic changes in the tilt angle histograms while lanosterol has essentially no effect. The addition of ergosterol has a slight influence at higher concentrations. Using the average tilt angle calculated from the single-molecule histograms, the order parameter S is calculated as a function of cholesterol and compared with previous studies.(8-10).