Vitreous fluorophotometry measurements taken from diabetic patients with different degrees of retinopathy and from normal volunteers were used to evaluate the integrity of the blood-ocular barrier. The contributions of lens and retinal fluorescence to the measured vitreous fluorescence were investigated. Lens autofluorescence was significantly higher for diabetics than for age-matched normals, and in age-matched diabetics the lens autofluorescence increased significantly with the progression of diabetic retinopathy. Natural ocular fluorescence was corrected for by subtracting baseline values from all vitreous measurements. Analysis of the baseline-corrected vitreous fluorescence values showed that both degree of retinopathy and age significantly affected the measured fluorescence. In age-matched groups, vitreous fluorescence values were greater in diabetics than in normals, and these values increased with the progression of retinopathy. Our results suggest that clinical vitreous fluorophotometry may be useful in evaluating the activity and predicting the progression of diabetic retinopathy.