Cholesterol-Induced Nanoscale Variations in the Thickness of Phospholipid Membranes

Graphene-induced energy transfer (GIET) is a recently developed fluorescence-spectroscopic technique that achieves subnanometric optical localization of fluorophores along the optical axis of a microscope. GIET is based on the near-field energy transfer from an optically excited fluorescent molecule to a single sheet of graphene. It has been successfully used for estimating interleaflet distances of single lipid bilayers and for investigating the membrane organization of living mitochondria. In this study, we use GIET to measure the cholesterol-induced subtle changes of membrane thickness at the nanoscale. We quantify membrane thickness variations in supported lipid bilayers (SLBs) as a function of lipid composition and increasing cholesterol content. Our findings demonstrate that GIET is an extremely sensitive tool for investigating nanometric structural changes in biomembranes.