Athletic animals and humans are commonly subjected to bone microdamage at subchondral (SCB) sites, found under joint cartilage, due to repetitive high-impact activity.
The accumulation of microdamage at focal joint sites can lead to fractures, severe joint disease and in some cases fatal bone injuries, therefore it is crucial to reliably locate and quantify the damage. In this paper we compared two incubation methods for staining bone specimens with lead uranyl acetate (LUA), i) a single-mix protocol in which the uranyl and lead reagents were combined and specimens incubated for 14 days; ii) a two-step method in which specimens were incubated sequentially in uranyl acetate and then lead acetate for seven days each. 3D images of the samples revealed that the two-step approach produced higher quantity and deeper labels of the microdamage.