This work describes composite structures composed of lipid bilayer or tetraetherlipid monolayer films attached to solid supports with associated crystalline bacterial cell surface layers (S-layers). The bilayer system was established by making use of the strong chemisorption of a first monolayer of thiolipids (1-octadecanethiol or 1,2-dimyristoyl-sn-glycero 3-phosphothioethanol) on gold and attaching a second monolayer of 1,2-dipalmitoyl-sn-3-phosphatidylethanolamine by the Langmuir Schaefer technique. The tetraetherlipid monolayer was composed of Glycerol-dialkyl-nonitol tetraetherlipid (GDNT). The monolayer of GDNT exhibits the thickness of a bilayer with hydrophilic headgroups on both sides and a hydrophobic inner part. Isolated S-layer protein from Bacillus sphaericus CCM2177, which was injected into the subphase of an LB-trough, recrystallized into a coherent monolayer at the solid supported phospholipid bilayer and at the tetraetherlipid monolayer. The composite lipid/S-layer structures were stable enough to allow lifting from the air-water interface, rinsing in water, and transfer into a scanning force microscope. (C) 1997 Academic Press.