Interfacial processes involving peripheral proteins depend on the composition and packing density of the interfacial lipid molecules. As a biological membrane model, lipid monolayers at the gas-liquid interface allow independent control of these parameters. However, measuring protein adsorption to monolayers has been difficult. To aid in this and other studies of the interfacial processes, we have developed an open, microfluidic flow cell with which surface physical properties can be controlled and monitored in well-defined lipid monolayers while varying aqueous-phase composition. Using this apparatus, we implement a recently described fluorescence method (Momsen, W. E.; Mizuno, N. K.; Lowe, M. E.; Brockman, H. L. Anal. Biochem. 2005, 346, 139-49) to characterize the adsorption/desorption of glucagon to 1,2-dioleoylsn-glycerol monolayers at 27 mN/m. Analysis of the data gives reasonable and self-consistent results for kinetic and thermodynamic constants. Varying the packing density of 1,2-dioleoyl-sn-glycerol does not alter the extent of glucagon adsorption, but comparable measurements with 1-steaoryl-2-oleoyl-sn-glycero-3-phosphocholine show a critical dependence. Because it allows a high degree of control of both lipid monolayer properties and aqueous-phase composition, this microfluidic flow cell should find wide applicability in many areas of research into interfacial processes.