Aims: To evaluate the safety and efficacy of transconjunctival 25 gauge vitrectomy under topical anesthesia. Settings and Design: A pilot study of consecutive cases which underwent 25 gauge vitrectomy under topical anesthesia. Materials and Methods: Seven eyes of 7 patients underwent 25 gauge vitrectomy under topical anesthesia with a pledget soaked in anesthetic, for vitreous hemorrhage (2 eyes), retained cortex (1 eye) and postoperative endophthalmitis (4 eyes). Subjective pain and discomfort were graded from 0 (no pain or discomfort) to 4 (severe pain and discomfort). Patients underwent an immediate postoperative assessment, followed by day one and one week postoperative evaluation. Results: All patients had grade 0 pain during the surgery. Five patients had grade 2 pain during the placement of the sclerotomies. None of the patients required any sedation during the procedure. No inadvertent eye movements were noted during surgery. Except one patient, none required postoperative analgesics. Five eyes had a favorable outcome. No eyes in this pilot study had any procedure-related complications. Conclusion: With appropriate case selection, topical anesthesia is a safe and effective alternative to infiltrative anesthesia for 25 gauge vitrectomy. A larger series of patients with a longer follow-up is required to validate the findings of this pilot study.