To characterize safety culture in emergency medical services (EMS), the authors modified a validated safety culture instrument, the Safety Attitudes Questionnaire (SAQ).The pilot instrument was administered to 3 EMS agencies in a large metropolitan area.The authors characterized safety culture across 6 domains: safety climate, teamwork climate, perceptions of management, job satisfaction, working conditions, and stress recognition.The feasibility of characterizing safety culture in EMS was evaluated by examining response rate, item missingness, EMS chief administrators’ perceptions of the EMS-SAQ, as well as psychometric properties.The results confirm feasibility with a high response rate, acceptable internal consistency, and model fit validity. However, some agencies voiced concerns about respondent burden and the wording and face validity of several EMS-SAQ items.Variation in safety culture scores across EMS agencies within a single geographic area, as well as variation across respondent characteristics, warrants further investigation.