| Abstract | The formation of domains in supported lipid membranes has been studied extensively as a model for the 2D organization of cell membranes. The compartmentalization of biological membranes to give domains such as cholesterol-rich rafts plays an important role in many biological processes. This article summarizes experiments from the author's laboratory in which a combination of atomic force microscopy and near-field scanning optical microscopy is used to probe phase separation in supported monolayers and bilayers as models for membrane rafts. These techniques are used to study binary and ternary lipid mixtures that have gel-phase or liquid-ordered domains that vary in size from tens of nanometers to tens of micrometers, surrounded by a fluid-disordered membrane. Examples are presented in which these models are used to investigate the distribution of glycolipid membrane raft markers and the preference for peptide and protein localization in ordered versus fluid membrane phases. Finally, the enzyme-mediated restructuring of membranes containing liquid-ordered domains provides an in vitro model for the coalescence of membrane rafts to give signaling platforms. Overall, the results demonstrate the importance of using techniques that can probe the nanoscale organization of membranes and of combining techniques that yield complementary information. Furthermore, the ability of supported lipid bilayers to model some aspects of membrane compartmentalization provides an important approach to understanding natural membranes. |
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