Osteomyelitis (OM), the bacterial infection of bone tissue, severely perturbs bone structure. The majority of OM is caused by Staphylococcus (S.) aureus, and even with proper treatment, OM has a high rate of recurrence and chronicity. While S. aureus has been shown to infect osteoblasts, persist there, and promote the release of pro-osteoclastogenic cytokines, it remains unclear whether osteoclasts (OCs) are also a target of intracellular infection. In this study, we examined the interaction between S. aureus and OCs. To test whether S. aureus could infect OCs in vivo, we injected GFP-expressing USA300 S. aureus subcutaneously over the periosteum of calvaria in TRAP-tdTomato OC reporter mice and visualized GFP+ S. aureus residing within OCs by confocal microscopy. We also found GFP+ S. aureus within OCs grown on bone slices and infected in vitro. Next, we employed an antibiotic survival assay during OC differentiation from bone marrow macrophages (BMMs) and found that the intracellular burden of S. aureus was increased at 18 hours post infection (hpi) in cells exposed to RANKL for at least 2 days (D2) compared to 1.5 hpi. Additionally, dividing bacteria were seen on transmission electron microscopy in OCs. In contrast, undifferentiated BMMs (D0) had fewer internal bacteria at 18 hpi. BMMs treated with interferon-γ also had fewer internal bacteria at 18 hpi compared to 1.5 hpi, whereas the internal bacterial load in BMMs treated with IL-4 was unchanged. Interestingly, flow cytometry of D2 BMMs infected with GFP+ bacteria showed a wide range of fluorescence. At 18 hpi, some cells maintained a bacterial load similar to 2 hpi, while others increased 100-1000 fold. In order to elucidate whether altered lysosomal localization could explain the ability of S. aureus to evade destruction in OCs, we utilized confocal microscopy with Lysotracker Red in OCs. We found a range of intracellular bacterial burdens that correlated inversely with S. aureus colocalization with lysosomes at 18 hpi. The heterogeneity of bacterial loads in both precursors and mature cells suggests that factors beyond the stage of OC differentiation are important. The ability of S. aureus to infect OCs and evade initial immune regulation may promote their proliferation at the periphery of lesions where OCs are most abundant. Better understanding of the interaction between OC lineage cells and S. aureus could point to host-targeted therapies that will impact progression or chronicity in OM.