Nowadays people are constantly under stress because of a fast-paced and demanding way of life which has serious consequences on their quality of life. Healthcare workers, especially those working in emergency care, experience a lot of stress due to long hours, large number of patients in need of immediate care and the emotional strain of the job. All of this is harmful to their mental health and can lead to burnout syndrome and affect patient care and healthcare as a whole. The purpose of this study is to compare factors affecting perceived stress in nursing staff based on whether they work in prehospital emergency care or hospital emergency department, as well as their level of education and whether they work in a small or a large town. The participants of this study were healthcare workers employed by the following institutions: Institute of Emergency Medicine of The City of Zagreb, University Hospital Centre Zagreb’s Department of Emergency Medicine, Institute of Emergency Medicine of Karlovac County and General Hospital Karlovac’s Department of Emergency Medicine. They were given two self-assessment questionnaires, one for stress level intensity and one for burnout syndrome intensity with additional sociodemographic questions. All of the research so far suggests a high rate of stress and burnout syndrome in healthcare, but with no significant difference between those employed in prehospital emergency care compared to those working in hospital emergency departments. The results of this study show a significantly higher overall stress experience in participants working in hospital emergency departments, and a higher degree of burnout syndrome in those with a higher level of education, again with no significant difference in stress levels depending on the size of the urban area nor in burnout syndrome rate depending on the place of employment or the size of the urban area.