To evaluate the perception of safety culture by health professionals who work with organ and tissue donation.A quantitative, descriptive study developed with 185 health professionals who act directly and indirectly with organ and tissue donation from two hospitals in the South of Brazil. The data collection was performed between January and July 2017 by using the Safety Attitudes Questionnaire. The analysis took place through descriptive statistics.The mean score of the domains evaluated ranged from 41.6, for Perception of management of the unit, and 80.9 for Job satisfaction. Positive perception of safety culture in this study was evidenced only for Job satisfaction with a score higher than 75.Of the six domains evaluated, only one had a positive score, evidencing the need to elaborate effective strategies for implanting safety culture in these institutions.