Atomic Force Spectroscopy (AFM) is a powerful technique that can be used to quantify the elastic properties of materials or to measure single molecule unfolding or unbinding interactions on the picoNewton scale. AFM probes are composed of fl exible, triangular or rectangular cantilevers with a sharp tip near the end of the cantilever. They can be manufactured from a variety of materials, but most AFM probes are made from silicon and/or silicon nitride (Si3N4) wafers using semiconductor- based etching processes. An AFM probe’s sensitivity, or spring constant (k), is the force required to bend the cantilever per unit distance (usually reported in Newtons/meter; N/m). It is an important factor in AFM probe behavior and performance for imaging applications and an essential parameter when attempting to quantify intra- or inter-molecular interactions or the compliance of materials with the AFM.