Maintaining precise temperature control is crucial for obtaining accurate isotherms. As temperature increases, deviations in the isotherm become more pronounced primarily due to the heightened molecular motion within the DPPC film complicates the replication of isotherms. This is obvious in Figure at temperature of 310 and 313 K. The general trend indicates that lift-off surface pressures shift towards higher molecular areas, while phase coexistence regions shift towards higher surface pressures. The collapsing surface pressure of the isotherm also shows more variation among the various sources as the temperature increases. To maximize isotherm reproducibility and accuracy, we performed surface pressure-area measurements under carefully controlled conditions, including precise temperature regulation (±0.5°C), controlled compression rates (10 mm/min), and standardized deposition protocols.