Abstract: Method of tuning parameter settings for performing acoustic scanning probe microscopy for subsurface imaging, scanning probe microscopy system, and computer program product. This document relates to a method of tuning a scanning probe microscopy system. The method comprises: a) applying an acoustic vibration signal comprising a first frequency and a second frequency to a sample; b) at a first position of the probe tip, sweeping the first frequency across a first frequency range, and obtaining a first signal; c) at a second position of the probe tip, sweeping the first frequency across at least said first frequency range, and obtaining a second signal; d) analyzing the first and second signals to obtain a difference characteristic dependent on the first frequency. The first and second position are selected such that a subsurface structure of the sample at the first and second position is different.