Effective Stiffness of Hydrated Atomic Force Microscopy Tips

When generating force curves with atomic force microscopy (AFM), the conventional assumption is that the silicon tip’s apex is infinitely stiffer than the force gradient acting between the apex and test object. Although true for measurements in vacuum or at long distances, we show this assumption fails badly at short distances in aqueous environments. In this case, the effective apex is an adsorbed water molecule, bound by a weak O–H···O–H H-bond. At short distances, the magnitude of the force gradient exceeds the stiffness of this bond. This causes conventional AFM measurements to be dominated by the mechanical H-bond stiffness, instead of the force gradient. Here, we introduce a new multifrequency technique that is able to measure the surface force gradient independently from the H-bond. We compare our results to conventional FM-AFM and show that due to the H-bond, FM-AFM can give extremely erroneous measurements and even the wrong force polarity.