A set-up enabling the recording of surface-enhanced Raman scattering (SERS) of phospholipid bilayers in contact with water is reported. Monolayers or bilayers of dimyristoyl-phosphatidylcholine (DMPC), palmitoyl-oleoyl-phosphatidylcholine (POPC) or palmitoyl-oleoyl-phosphatidylethanolamine (POPE) and cholesterol were transferred by the Langmuir-Blodgett technique onto a high index rutile prism on which a silver coating (15 nm thick) had been deposited. The bilayers were enclosed in a water-tight box. The prism is illuminated with a laser beam under an angle greater than the limiting reflection angle. SERS spectra of monolayer or bilayer films were compared with the Raman spectrum of anhydrous molecules. An enhancement in the relative intensity of bands corresponding to the vibrations of the polar head group was observed. In the 2750-3050 cm-1 region the monolayer spectra of POPC and DMPC were different from those of the bilayers; this difference was assigned to the mobility of the choline CH3 groups, near the silver coating. No differences appear between SERS spectra of POPC-cholesterol bilayers and those of pure POPC bilayers.