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Th aim of this study is to explore the diagnostic accuracy of smartphone-based D-EYE ophthalmoscopy in the diagnosis of optic nerve head (ONH) abnormalities and screening for glaucoma. This is a prospective clinical-based validation study performed on 90 patients recruited from a tertiary teaching hospital. Patients underwent dilated fundus examination by slit-lamp and smartphone-based D-EYE fundoscopy operated by two experienced ophthalmologists. The diagnostic accuracy of smartphone-based D-EYE ophthalmoscopy of normal vertical cup-to