BACKGROUND & AIMS: Omega 3 and omega 6 polyunsaturated fatty acids have been related to many diseases, especially to cardiovascular diseases. We aimed to assess the association between the intake of omega 3, omega 6 fatty acids and their ratio and the incidence of glaucoma in this prospective cohort study. METHODS: We followed-up 17,128 participants initially free of glaucoma for a median time of 8.2 years. Validated data of diet were collected at baseline with a 136-item semi-quantitative food-frequency questionnaire and information of new diagnosis of glaucoma in biennial follow-up questionnaires. Multivariable Cox regression models were fit to assess the relationship between omega 3, omega 6 and their ratio and the incidence of glaucoma in the total sample and in participants aged ≥40 years at baseline. RESULTS: During a median follow-up time of 8.2 years, 156 new cases of glaucoma were identified. No significant association was observed for the omega 3 or the omega 6 intake and the risk of glaucoma. Participants in the highest quintile of omega 3:6 ratio intake had a significantly higher risk of glaucoma than participants in the lowest quintile (hazard ratio (HR): 1.91 [95%CI: 1.05-3.46], p for trend 0.03). The association became stronger (HR for the comparison of the 5th versus the 1st quintile: 2.43 [95%CI: 1.17-5.03], p for trend 0.02) when we considered only those participants who were ≥40 years old. CONCLUSIONS: Our data suggest an association between omega 3:6 ratio intake and incident glaucoma in our cohort, especially among older participants.