Membrane lipids are structurally diverse in ways that far exceed the role envisioned by Singer and Nicholson of simply providing a fluid bilayer matrix in which proteins reside. Current models of lipid organization in membranes postulate that lipid structural diversity enables nonrandom lipid mixing in each leaflet of the bilayer, resulting in regions with special physical and functional properties, i.e., microdomains. Central to understanding the tendencies of membrane lipids to mix nonrandomly in biomembranes is the identification and evaluation of structural features that control membrane lipid lateral mixing interactions in simple model membranes. The surface balance provides a means to evaluate the lateral interactions among different lipids at a most fundamental level-mixed in binary/ternary combinations that self-assemble at the air-water interface as monomolecular films, i.e., monolayers. Analysis of surface pressure and interfacial potential as a function of average cross-sectional molecular area provide insights into hydrocarbon chain ordering, lateral compressibility/elasticity, and dipole effects under various conditions including those that approximate one leaflet of a bilayer. Although elegantly simple in principle, effective use of the surface balance requires proper attention to various experimental parameters, which are described herein. Adequate attention to these experimental parameters ensures that meaningful insights are obtained into the lipid lateral interactions and enables lipid monolayers to serve as a basic platform for use with other investigative approaches.