Higher harmonic atomic force microscopy: imaging of biological membranes in liquid.

The contribution of higher harmonics to the movement of a dynamic force microscope cantilever interacting with a sample in liquid was investigated. The amplitude of the second harmonic has been found to be an order of magnitude higher in liquid than in air, reflecting an increased sensitivity to local variations in elasticity and interaction geometries. A theoretical model of the tip-sample interactions in liquid was introduced and shown to be consistent with experimental findings. Second harmonic amplitude images were recorded on soft biological samples yielding a lateral resolution of approximately 0.5 nm.