The use of biopolymers in nanotechnologic applications, e.g. as coatings, is often limited by their suboptimal nanomechanical properties. Here we used an atomic force microscopy (AFM) based approach to study the elastic properties and thickness of surface supported DOPC membranes containing varying amounts of bolalipids (C(20)BAS) at the nanoscale. We found that increasing amounts of bolalipids led to higher stability and decreasing thickness of the bilayer membrane. A combination with fluorescence microscopy further allowed us to investigate phase separation on several length scales. We anticipate that similar characterizations will be helpful to optimize multi-component biopolymer coatings. (C) 2015 Elsevier Ltd. All rights reserved.