In order to increase limited organ donations, public awareness must be raised. It is also essential that health personnel have enough knowledge about organ donation. This research aims at determining health personnel knowledge and attitudes about organ donation in a tertiary hospital with an organ transplant center.This prospective, cross-sectional research was conducted between January 10, 2017 and March 1, 2017, in Gülhane Education and Research Hospital in Ankara, Turkey. A survey including 20 questions about sociodemographic characteristics and organ donation was used. Twelve hundred surveys were distributed and 1012 of them (84.3%) were analyzed.Age range of the participants was 18 to 63 years. Of the 1012 participants, 241 (23.8%) were physicians, 202 (20%) were nurses, 220 (21.7%) were health technicians, and 349 (34.5%) were nonclinical staff. Of all participants, 603 (59.6%) had enough information about organ donation, 431 (42.6%) chose media and internet as information sources about organ donation. It was found that 534 (52.8%) of health personnel were volunteers for organ donation; however, only 169 (16.7%) of them had an organ donation card.More than half of health personnel were volunteers themselves for organ donation and had enough knowledge about this subject. Physicians were more positive about organ donation than other health personnel.