BACKGROUND AND OBJECTIVE: To assess the surgical outcomes of the use of tissue glue to close sclerotomy sites when required and the views of ultrasound biomicroscopy of the sclerotomy sites in 23- and 25-gauge vitrectomy systems. PATIENTS AND METHODS: A 25-gauge transconjunctival sutureless vitrectomy was performed in 38 eyes and a 23-gauge transconjunctival sutureless vitrectomy was performed in 46 eyes for various vitreoretinal diseases. Wound leakage occurred at the sclerotomy sites at the end of the surgery in 6 eyes with 23-gauge transconjunctival sutureless vitrectomy and 7 eyes with 25-gauge transconjunctival sutureless vitrectomy. The sclerotomy sites were closed by using tissue glue to prevent wound leakage and evaluated with ultrasound biomicroscopy postoperatively. RESULTS: No wound leakage was observed at the end of the surgical procedure or during the follow-up period. Abnormal fibrous ingrowth was not detected at the sclerotomy sites by means of ultrasound biomicroscopy. CONCLUSION: The results demonstrated the efficacy of tissue glue for closing site ports when wound leakage is observed in transconjunctival sutureless vitreoretinal surgery.