Staphylococcus aureus is an important human pathogen, capable of infecting nearly all organ systems in the body. The ability of S. aureus to infect such a broad spectrum of host tissues requires flexible metabolic programs to enable proliferation within diverse environments. The skeleton is an exceptionally common site of invasive staphylococcal infection, and bone infections are notoriously recalcitrant to antibiotic therapy due to limited antibiotic penetrance into damaged bone and rapid induction of antimicrobial tolerance in invading bacteria. This seminar will highlight: 1) mechanisms that drive the pathogenesis and antibiotic recalcitrance of osteomyelitis, with a focus on bacterial metabolism, virulence regulation, and antibacterial immune defenses; 2) novel molecular imaging approaches to query spatially refined host-pathogen biology; and 3) ongoing efforts to understand the role of osteo-immunologic crosstalk in infectious disease.