Abstract

Much of lipid raft properties can be inferred from phase behavior of multicomponent lipid membranes. We use liquid compatible atomic force microscopy (AFM) to study a three-component system composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), egg sphingomyelin (eSM), and cholesterol. Specifically, we obtain macroscopic and nanoscopic heterogeneous structures in a broad compositional space of DOPC/eSM/cholesterol (23 °C). In the macroscopic liquid coexisting region, we quantify area fraction of the coexisting phases and determine a set of thermodynamic tie-lines. When lipid compositions are near the critical point, we obtain fluctuation-like nanoscopic structures. We also use AFM height images to explore the hypothetical three-phase coexisting region. Finally, we use fluorescence microscopy to compare the phase behavior from our AFM measurements to that in free-floating giant unilamellar vesicles (GUVs). Our results highlight the role of lipid composition in mediating lipid domain formation and stability.

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