Surface Pressure-Area Isotherm Collection. Surface pressure-area isotherms were collected with a Biolin Nima Langmuir medium trough (max area: 243 cm 2 ) at the air-subphase interface. The aqueous subphase consisted of ultrapure water (18.2 MΩ•cm resistivity, TOC < 5 ppb) obtained from a Millipore purification system, with pH and surface tension measured before each experiment to verify subphase quality, whose temperature was precisely controlled using a water circulating bath with an accuracy of ± 0.5 o C. A precisely measured aliquot (30 µL) of DPPC solution (1 mg/mL in HPLC-grade chloroform), doped with 0.5 wt% TR-DHPE as a fluorescent probe, was carefully deposited onto the aqueous subphase using a gas-tight Hamilton microsyringe to ensure reproducible spreading conditions. Stock solutions were prepared in amber glass vials, purged with nitrogen to prevent lipid oxidation, and stored at -20°C; working solutions were equilibrated to room temperature for 30 minutes prior to monolayer deposition to prevent temperature-induced spreading artifacts. The compression rate was optimized at 10 mm/min based on preliminary studies showing this speed allows sufficient time for molecular reorganization while minimizing film loss through subphase dissolution and maintaining experimental feasibility; faster rates led to artifactual smoothing of phase transitions while slower rates increased susceptibility to environmental perturbations. Each isotherm measurement was performed in triplicate using fresh monolayers and subphases, with statistical analysis of key transition points (lift-off area, collapse pressure, phase transition plateau) showing coefficients of variation < 5% across independent measurements.