Colloidal interaction forces between a silica particle and a solid-supported Langmuir-Schaefer phospholipid bilayer were directly measured using a gradient optical trap and evanescent wave light scattering. A small custom-built Langmuir trough was integrated with an optical trapping microscope to allow force measurements on a single particle within the subphase of the trough after the dip of the substrate was completed. The novel method allows the force measurements to be conducted without transferring the substratum across an air/water interface. The fluctuating particle position near the bilayer was tracked by evanescent wave light scattering to determine the deflection due to surface forces, and the relaxation time of particle fluctuations was measured to simultaneously determine the viscous forces. Measured equilibrium and viscous force-distance profiles of silica microspheres with diameters of I and 5 pm on bilayers of dipalmitoyl phosphatidyl choline (DPPC) were markedly different than force-distance on bare mica and DPPC monolayers under the same electrolyte conditions. (c) 2006 Elsevier Inc. All rights reserved.