Osteomyelitis in children can produce significant debilitation and broadly includes infectious (acute and chronic) and non-infectious osteomyelitis, including chronic recurrent multifocal osteomyelitis (CRMO). Biopsy is often performed to aid in diagnosis and to identify the pathogen, frequently in cases wherein best management, based on clinical presentation, is ambiguous. The aim of our study was to characterize the yield and assess efficacy of image-guided bone biopsy for pediatric osteomyelitis. This was a retrospective review of patients with suspected osteomyelitis based on clinical data and imaging who underwent percutaneous bone biopsy in interventional radiology over a 10-year period at a tertiary pediatric hospital. Review parameters included procedural technical data and clinical data, including details of clinical presentation and microbiology and pathology results. 40 patients (mean age 9.5y, range 0.4-25.8y) underwent biopsy. Fluoroscopy was used in 29 cases, with rotational CT in 16 cases. Of these 16, Syngo iGuide guidance software was used in 9 cases, and MRI overlay was used in 4. CT guidance was used in 9. Additional US guidance for adjacent soft tissue component was used in 17 cases. Mean procedure time was 85 minutes, and mean fluoroscopy time was 3.6 min. A 16G bone biopsy cannula was utilized in 34 cases; a 13G cannula in 5; and an 11G in 1 case. The mean number of bone cores obtained was 3.6. Biopsy culture yielded a pathogen in 4 of 40 cases, and culture results did not show significant correlation with lesion location, fever, leukocytosis/blood culture, ESR/CRP, number of cores, instrument gauge, or aspiration of marrow. There was pathology adequacy in 100%. Based on culture and pathology results, 22 patients were diagnosed with infectious osteomyelitis (13 chronic and 9 acute) and were placed on antibiotic therapy. Fifteen patients were diagnosed with CRMO, based on pathology result. Although culture yield of bone biopsy was low, pathology results showed adequacy and aided in management of patients with suspected osteomyelitis. Results support bone biopsy as a safe and effective means to aid in management of pediatric osteomyelitis.