Abstract Mixtures of 1,2 Distearoyl-sn-glycero-3-phosphocholine (DSPC) and polyoxyethylene-40-stearate (PEG40St) were prepared at different molar ratios and their miscibility were investigated using Langmuir isotherms. Pure DSPC monolayer exhibited a liquid-condensed (LC) phase whereas PEG40St monolayer exhibited liquid-expanded (LE) phase at the air-water interface at 22 ± 2 °C. At the collapse pressure of 33 mN/m, the PEG40St mean molecular area was calculated to be 28 A2/molecule for 9:1 composition and 50 A2/molecule for 5:5 composition, showing an increasing trend with the emulsifier content. A quantification method was developed to estimate the squeeze out amount of PEG40St from Langmuir isotherms of the DSPC/PEG40St mixtures at different molar ratios. Almost 93%, 82%, and 53% of PEG40St displaced for the 9:1, 7:3, and 5:5 mixtures, respectively, at the end of the first collapse plateau and showed a decreasing trend with the PEG40St content. Remaining PEG40St squeezed out at the end of the second collapse plateau, where 20% of PEG40St still contained within the 5:5 composition. It was concluded that increasing PEG40St content would be advantageous to design more stable lipid based microbubbles.