This study investigated the impact of working for public versus private ambulance services in Turkey and elaborated on predictors of mental, physical, and emotional well-being in emergency medical technicians (EMT-Bs). In this observational cross-sectional study, an 81-question self-report survey was used to gather data about employee demographics, socioeconomic status, educational background, working conditions, and occupational health and workplace safety (OHS), followed by the 12-item General Health Questionnaire (GHQ-12), the Work-Related Strain Inventory (WRSI), and the Maslach Burnout Inventory (MBI) with three subscales: Emotional Exhaustion (MBI-EE), Depersonalization (MBI-DP), and Diminished Personal Accomplishment (MBI-PA). In 2011, 1,038 EMT-Bs worked for publicly operated and 483 EMT-Bs worked for privately owned ambulance services in Istanbul, Turkey, of which 606 (58.4%) and 236 (48.9%) participated in the study (overall participation rate = 55.4%), respectively. On all scales, differences between total mean scores in both sectors were statistically insignificant (p > .05). In the public sector, work locations, false accusations, occupational injuries and diseases, work-related permanent disabilities, and organizational support were found to significantly influence self-reported perceptions of well-being (p < .05). In the private sector, commute time to and from work (p < .05), false accusations (p < .05), vocational training and education (p < .05), informed career choices (p < .05), and work-related permanent disabilities (p < .05) were found to significantly influence self-reported perceptions of well-being. EMT-Bs were asked about aspects of their working lives that need improvement; priority expectations in the public and private sectors were higher earnings (17.5%; 16.7%) and better social opportunities (17.4%; 16.8%). Working conditions, vocational training, and OHS emerged as topics that merit priority attention.