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To assess the internal reflectivity of the retinal nerve fiber layer in normal, ocular hypertensive, and glaucomatous eyes using optical coherence tomography. Methods: All patients underwent complete ophthalmic examination and achromatic automated perimetry. Intraocular pressure was 21 mm Hg or less for low-tension glaucoma and normal eyes and at least 25 mm Hg on 2 separate occasions in ocular hypertensive and high-tension glaucoma eyes. All glaucomatous eyes had characteristic glaucomatous optic neuropathy and associated achromatic automated perimetry defect. Relative retinal nerve fiber layer internal reflectivity was measured on optical coherence tomography images using a software program of our own design. Results: We enrolled 98 eyes (19 normal, 34 ocular hypertensive, 17 high-tension glaucoma, and 28 lowtension glaucoma). Relative internal reflectivity was less in eyes with glaucoma than in normal (PϽ.001, t test) and ocular hypertensive eyes (PϽ.001, t test). There was no difference in relative internal reflectivity between normal and ocular hypertensive eyes (P=.32) and between eyes with high-tension glaucoma and low-tension glaucoma (P = .43). Internal reflectivity correlated with mean deviation on achromated automatic perimetry (r 2 = 0.49, PϽ.001, quadratic regression analysis). Conclusion: Relative retinal nerve fiber layer internal reflectivity may provide useful information about the extent of retinal nerve fiber layer injury in glaucoma.