Dynamic acousto-elastic testing (DAET) is based on the coupling of a lowfrequency (LF) acoustic wave and high-frequency ultrasound (US) pulses (probing wave). It was developed to measure US viscoelastic and dissipative non-linearity in trabecular bone. It is well known that this complex biphasic medium contains microdamage, even when tissues are healthy. The purpose of the present study was to assess the sensitivity of DAET to monitor microdamage in human calcanei. Three protocols were therefore performed to investigate the regional heterogeneity of the calcaneus, the correlation between DAET measurements and microdamage revealed by histology, and DAET sensitivity to mechanically induced fatigue microdamage. The non-linear elastic parameter b was computed for all these protocols. The study demonstrated the presence of high viscoelastic and dissipative non-linearity only in the region of the calcaneus close to the anterior talocalcaneal articulation (region of high bone density). Protocols 1 and 2 also showed that most unsorted calcanei did not naturally exhibit high non-linearity, which is correlated with a low level of microcracks. Nevertheless, when microdamage was actually present, high levels of US non-linearity were always found, with characteristic non-linear signatures such as hysteresis and tension/compression asymmetry. Finally, protocol 3 demonstrated the high sensitivity of DAET measurement to fatigue-induced microdamage.