The use of clinical practice guidelines (CPGs) saves time and provides the highest level of evidence, allowing busy ophthalmologists to make the best clinical decisions in a short amount of time. The aim of this study is to evaluate Iranian ophthalmologists’ knowledge and attitudes toward evidence-based medicine (EBM), and knowledge of CPGs. We performed a cross-sectional survey during the Annual Congress of the Iranian Society of Ophthalmology (November 2011). We administered 2 questionnaires regarding ophthalmologists’ attitudes towards EBM and self-reported clinical practices to a self-selected sample of ophthalmologists who volunteered to be surveyed. We employed descriptive statistics and unadjusted associations to describe the results. Of 556 attendees, 100 ophthalmologists responded to the first questionnaire, and 60% of them answered the second questionnaire. Although 83% had heard about EBM, only 11% knew what EBM was. Ninety-five percent of respondents believed in evidence-based ophthalmology training. Few understood EBM terminology. More than 60% of ophthalmologists surveyed had no familiarity with the Cochrane Library, MDConsult or Turning Research Into Practice (TRIP databases. Of the ophthalmologists surveyed, 26% implemented guidelines for problem solving, and 59% utilised the internet for finding clinical guidelines. The mean proportions of correct response to evidence-based cataract and glaucoma management questions were 68 and 56, respectively. Iranian ophthalmologists had a fair to poor knowledge but good attitudes towards EBM. Improving evidence-based knowledge and attitudes may be a component of improving Iranian ophthalmologists’ clinical practices.