In vivo measurement of metabolic changes in diabetic cornea and lens were performed using redox fluorometry in nonobese diabetic (NOD) mice. Autofluorescence from reduced pyridine nucleotides (PN) and oxidized flavoproteins (Fp) were measured, and the PN/Fp ratio was used as a tissue metabolism indicator. The PN/Fp ratios were significantly higher in the diabetic corneal endothelium. Morphometric analysis of the corneal endothelium using specular microscopy revealed no significant differences between the two groups. These results indicate that redox fluorometry is able to detect early metabolic changes in the corneal endothelium and lens epithelium, which are induced by diabetes mellitus. Activation of the polyol pathway may be responsible for the change. Corneal epithelia may be less susceptible to diabetic changes than the corneal endothelium and lens epithelium.