Abstract Background: This study aims to investigate the effects of tropicamide 1% on the main numerical parameters of anterior segment with the Pentacam rotating Scheimpflug camera in healthy individuals. Methods: Pentacam measurements of 50 (25 female and 25 male) healthy individuals before and after 25 min of tropicamide 1% instillation were performed and compared. For comparison of the test parameters, paired t -test was used in the groups. Results: Mean age in the study group was 28.40±4.58 years (range 18–35 years). Measurements between 2 sessions were significantly different for the parameters of anterior chamber depth (ACD) and anterior chamber volume (ACV) ( P <0.05). Conclusions: Tropicamide 1% instillation leads to a significant increment in ACD and ACV measurements. Effects of tropicamide on these parameters for Pentacam measurements should be considered to ensure exact clinical interpretation in ophthalmologic practice.