Stress fractures are frequently observed in physically active populations, and they are believed to be associated with microcrack accumulation. There are not many tools for real-time monitoring of microdamage formation during fatigue of bone, in vivo or in vitro. Acoustic emission (AE) based detection of stress waves resulting from microdamage formation is a promising method to assess the rate and energetics of microdamage formation during fatigue. The current study aims to assess the time history of the occurrence of AE events during fatigue loading of human tibial cortical bone and to determine the associations between AE variables (energy content of waves, number of AE waveforms, etc.), fatigue life, and bone ash content. Fatigue test specimens were prepared from the distal diaphysis of human tibial cortical bone (N = 32, 22 to 52 years old, male and female). The initiation of acoustic emissions was concomitant with the nonlinear increase in sample compliance and the cumulative number of AE events increased asymptotically in the prefailure period. The results demonstrated that AE method was able to predict the onset of failure by 95% of the fatigue life for the majority of the samples. The variation in the number of emissions until failure ranged from 6 to 1861 implying a large variation in crack activity between different samples. The results also revealed that microdamage evolution was a function of the level of tissue mineralization such that more mineralized bone matrix failed with fewer crack events with higher energy whereas less mineralized tissue generated more emissions with lower energy. In conclusion, acoustic emission based surveillance during fatigue of cortical bone demonstrates a large scatter, where some bones fail with substantial crack activity and a minority of samples fail without significant amount of crack formation.