Several methods of measuring the line tension between phase-separated liquid-ordered–liquid -disordered domains in phospholipid–cholesterol systems have been proposed. These experimental techniques are typically internally self-consistent, but the measured line tension values vary widely among these techniques. To date, no measurement of line tension has utilized multiple experimental techniques to look at the same monolayer system. Here we compare two nonperturbative methods, Fourier analysis of boundary fluctuations (BA) and one proposed by Israelachvili involving the analysis of domain size distributions (SD), to extract the line tension in a 70 mol % DMPC/30 mol % dihydrocholesterol (DChol) mixture as a function of surface pressure. We show that BA predicts the expected variation in line tension measurements consistent with the theoretical critical exponent whereas SD does not. From this comparison, we conclude that the size distribution of monolayer domains is metastable and primarily determined by the kinetics of domain nucleation and subsequent aging.

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