Phase Transitions in a Single Supported Phospholipid Bilayer: Real-Time Determination by Neutron Reflectometry.

Time- and temperature-resolved neutron reflectometry allowed us to perform the real-time characterization of the structural changes taking place across phase transitions in solid supported-lipid bilayers (SLBs). We identified the presence of an isothermal phase transition, characterized by a symmetrical rearrangement of lipid molecules in both bilayer leaflets, followed by the main thermotropic phase transition, and characterized by an independent melting of the two leaflets. We demonstrated that the presence of a substrate increases the enthalpy of melting by the same amount for both SLB leaflets with respect to the values reported for freestanding bilayers. These results are highly relevant for the further understanding of cooperative structural dynamics in SLBs and for the investigation of thermally activated processes such as the transmembrane lipid translocation (flip flop).