AIM: To report the cytology results of 25-gauge transconjunctival (25G-TSV) diagnostic vitrectomy in cases suspicious for intraocular lymphoma (IOL), and compare the results to those reported in the literature.METHODS: Clinical and cytopathological records of 18 vitreous biopsy specimens obtained via 25G-TSV diagnostic vitrectomy in 12 patients suspicious for IOL were reviewed retrospectively. A review of the literature in regards to the diagnostic yields of vitreous specimens obtained via 25-gauge and 20-gauge diagnostic vitrectomy in suspected cases of IOL was performed.RESULTS: Eighteen eyes from 12 patients with clinical suspicion of IOL underwent diagnostic 25G-TSV. The cytopathological investigations demonstrated IOL in 15 eyes (83.3%). Vitreous analysis was non-diagnostic in 3 eyes (16.7%).CONCLUSION: Twenty-five-gauge diagnostic vitrectomy yields adequate sample for cytological evaluation of the vitreous in cases suspicious for IOL. The diagnostic results of the 25G-TSV in the current study are superior to those reported for 20-gauge vitrectomy but equivalent to those reported for 25G-TSV in the published literature.