Bicelles have been intensively studied for use as drug delivery carriers and in biological studies, but their preparation with low-cost materials and via a simple process would allow their use for other purposes as well. Herein, bicelles were prepared through a semi-spontaneous method using a mixture of hydrogenated soybean lecithin (SL) and a nonionic surfactant, polyoxyethylene cholesteryl ether (ChEO10), and then we investigated the effect of composition and temperature on the structure of bicelles, which is important to design tailored systems. As the fraction of ChEO10 ( X C) was increased, a bimodal particle size distribution with a small particle size of several tens of nanometers and a large particle size of several hundred nanometers was obtained, and only small particles were observed when X C ≥ 0.6, suggesting the formation of significant structure transition (liposomes to bicelles). The small-angle neutron scattering (SANS) spectrum for these particles fitted a core-shell bicelle model, providing further evidence of bicelle formation. A transition from a monomodal to a bimodal size distribution occurred as the temperature was increased, with this transition taking place at lower temperatures when higher SL-ChEO10 concentrations were used. SANS showed that this temperature-dependent size change was reversible, suggesting the SL-ChEO10 bicelles were stable against temperature, hence making them suitable for several applications.