In this paper, we examined the effect of Laminaria japonica polysaccharides (LJP) on cationic 1,2-Dioleoyl-3-Trimethylammonium-Propane (DOTAP) and anionic 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-1-glycerol] (DPPG) monolayers at the air-water interface by the pressure-area isotherms (π-A), adsorption curves (π-t) and morphology measurements with atomic force microscopy (AFM) technique. The π-A curves revealed that the isotherms shifted to larger mean molecular area with progressive addition of LJP into subphase for both DOTAP and DPPG monolayers. And the compression modulus Cs−1 obtained from π-A curves showed that the elasticity of the films decreased with the addition of LJP. Adsorption curves were measured at the surface pressure of 10 and 20mN/m, which were fitted by the adsorption kinetics equation. It revealed that DOTAP monolayer changed into a mixed film with the insertion of polysaccharides molecules. However, there was no significant effect on the surface pressure for DPPG monolayer. Besides, surface morphology was observed by AFM, which was consistent with the results of fitted adsorption curves.