OBJECTIVE To understand the impact of storage temperature on recovery of Staphylococcus aureus on sampling swabs. Staphylococcus aureus is a common cause of skin and soft tissue infections, but also causes a variety of life-threatening diseases. With a large pool of asymptomatic carriers and transmission that can occur even through indirect contact, mitigation efforts have had limited success. Swab sampling, followed by culturing, is a cornerstone of epidemiological studies, however, S. aureus viability on swabs stored at different temperatures has not been characterized. RESULTS We determined survival rates on swabs stored at five different temperatures. Samples stored at -70°C had no decay over time while samples stored at higher temperatures showed an exponential decay in viability. Mortality rates were greatest for swabs stored at 37°C. Survival at intermediate temperatures (-20 to 20·5°C) did not differ significantly, however, we observed more variation at higher temperatures. CONCLUSIONS To maximize recovery of S. aureus cells, samples should be stored at -70°C or processed for culturing without delay. CONCLUSIONS Epidemiological studies of bacterial diseases are typically limited to determination of pathogen presence/absence, yet quantitative assessments of pathogen load and genetic diversity can provide insights into disease progression and severity, likelihood of transmission and adaptive evolutionary potential. For studies of S. aureus where time or access to a microbiology laboratory may delay culturing, deep freezing or timely culturing will maximize the degree to which sampling results reflect source status.