What do we really measure in AFM punch-through experiments on supported lipid bilayers?

Nowadays, there is much experimental evidence that the mechanical properties of biological membranes affect membrane protein functions. A very convenient technique to study these properties on a spatial scale relevant to that of single proteins is represented by Atomic Force Spectroscopy (AFS). In this study we measured the force the AFM tip has to apply on a supported lipid bilayer to punch-through it as a function of different environmental parameters. We observed that this force is reduced when the lipid bilayer is in its phase transition region. We interpreted our results on the basis of thermodynamical considerations and we stressed their biological relevance. In particular, the reduced punch-through force in the phase transition region could be relevant for the function of membrane proteins which operates by conformational changes at the protein/lipid interface. We also suggest that the presence of a transmembrane voltage drop can affect the measured punch-through force.