PURPOSE: To study 25-gauge sclerotomy healing process in vivo with ultrasound biomicroscopy (UBM) in direct and oblique incisions. DESIGN: Prospective interventional case series report. METHODS: At our institution, we performed UBM studies on 53 sclerotomies during the first 30 days after 25-gauge vitrectomy, looking for conjunctival bleb development, sclerotomy healing signs, and vitreous incarceration in the wound. RESULTS: Echographical healing signs were completed in 77% of patients by day 15 with no differences between direct and oblique sclerotomies. By day 30, all but one sclerotomy were closed. Conjunctival blebs developed over 64% of direct sclerotomies, and 25% of oblique (P = .0059), but all resolved spontaneously by day 15. Vitreous incarceration appeared in 72% of sclerotomies. CONCLUSIONS: Twenty,five gauge sclerotomies heal by day 15 in most cases with no difference between direct and oblique sclerotomy construction. Conjunctival blebs developed more frequently over direct than oblique sclerotomies.