AIM: To assess central corneal thickness (CCT) changes measured with Scheimpflug device following instillation of 2% fluorescein in normal subjects. METHODS: This was a prospective randomized study of 60 hospital volunteers. After baseline CCT measurements of both eyes of 40 subjects were obtained using Scheimpflug system, a drop of preservative-free 2% fluorescein, was instilled in one eye and in other eye, one drop of normal saline (control). Measurements were repeated after 1, 2, 5, 10, 20, 30, 40, 50 and 60min (continuous assessment group). Twenty subjects had baseline CCT taken, then fluorescein was instilled in one eye and measurements were taken at 1min. Ten eyes had saline rinse after 1min and 10 other eyes did not, measurements were repeated at 2min (eye rinse group). RESULTS: The mean baseline CCT for continuous assessment group was 546.2 ±32.1 μm (range, 489.0-606.0), control eyes was 546.6±30.7 μm (range, 489.0-602.0). At 1min after fluorescein instillation, CCT significantly increased by 37.0±34.0 μm (P<0.001), then decreased gradually, reaching baseline at 60min. CCT variations were not significant in control group (P>0.05). For eye rinse group, CCT mean differences between baseline and 2min were 18.2 μm (95 % CI: -54.7 to 18.3) with rinse and 26.5 μm (95% CI: -62.9 to 9.9) without rinse; paired sample tests were not significant (P>0.05). CONCLUSION: The presence of fluorescein increased CCT value to a clinically relevant level of 6.8%. Eye rinse did not significantly reduce the effect at 2min post fluorescein timepoint.