Fluorescence Microscopy to Study Domains in Supported Lipid Bilayers

Fluorescence microscopy of model membranes is a powerful tool for identifying the nature and extent of coexisting phases in biologically relevant lipid mixtures. Planar supported bilayers offer the advantage over spherical model membranes in that both overall composition and lipid asymmetry can be controlled. In addition, the membrane can be easily accessed by perfusion of soluble components. Here, the necessary techniques for reconstituting bilayers of complex composition and phase behavior in planar systems are outlined. Effective methods for the formation of both symmetric and asymmetric bilayers are described. Considerations that must be taken into account in choosing suitable lipid compositions, effective fluorescent lipid dyes, and adequate microscope setups are discussed.