In this work, the Langmuir monolayers were used as a model for the analysis of the influence of cholesterol on 1,2-distearoyl-sn-glycero-3-phosphocholine - DSPC and stearoyl sphingomyelin - SSM, as well as their equimolar mixture. The aim of these studies was to compare the affinity of cholesterol to sphingomyelin and phosphatidylcholine and discuss the effectiveness of cholesterol packing with these phospholipids. The experiments involved the registration of the surface pressure-area isotherms combined with the application of Brewster angle microscopy (BAM) images and grazing incidence X-ray diffraction methods. We have performed a thorough analysis of the properties of both one-component DSPC and SSM films as well as their 1:1 mixture. Next, the effect of cholesterol on these systems was verified based on the results for 2:1 SSM/Chol, 2:1 DSPC/Chol, and 1:1:1 DSPC/SSM/Chol mixtures. It was found that both phospholipids form highly condensed monolayers, however, they differ in the orientation of acyl chains, namely the acyl chains are more tilted in DSPC film as compared to SSM monolayer as well as DSPC/SSM mixture. Furthermore, the area contraction provoked by the addition of cholesterol was found to be more pronounced for DSPC monolayer than in DSPC/SSM and SSM films. However, all the collected results allow one to postulate that the ability of cholesterol to form ordered domains with DSPC and SSM is similar and is predominantly driven by hydrophobic forces between molecules. The differences in the area condensation induced by cholesterol on the studied phospholipids films results from differences in molecular organization of pure phospholipids films rather than specific cholesterol-phospholipid interactions. (C) 2013 Elsevier Inc. All rights reserved.