OBJECTIVE: To describe the technique and evaluate short- and long-term outcomes in female dogs after endoscopic-guided laser ablation (ELA) of various vestibulovaginal septal remnants (VVSRs). DESIGN: Retrospective case series. ANIMALS: 36 dogs. PROCEDURES: Medical records of dogs with VVSRs that underwent ELA were retrospectively reviewed. All patients underwent complete cystourethrovaginoscopy for diagnosis and treatment. Endoscopic-guided laser ablation (with a holmium:yttrium-aluminum-garnet or diode laser) was used to transect the vaginal membrane. Patients with intramural ectopic ureters were concurrently treated with ELA of their ectopic ureters. Endoscopy was repeated 6 to 8 weeks after ELA of vaginal remnants in some patients, and the procedure sites were reassessed. RESULTS: 36 female dogs with persistent paramesonephric septal remnants (n = 19), vaginal septa (11), or dual vaginas (6) were included. Twenty-six dogs had urinary incontinence, 2 had recurrent UTIs, and 8 had both. Thirty of 36 (83%) dogs had concurrent ectopic ureters. Endoscopic-guided laser ablation was performed with holmium:yttrium-aluminum-garnet and diode lasers in 8 and 28 dogs, respectively. Five dogs had mild postoperative dysuria for < 24 hours. One patient developed a complication involving inadvertent laser perforation of the vaginal wall. There were no negative effects from this event, and the perforation was fully healed within 8 weeks. At the time of follow-up, all defects were fully healed with no sign of recurrence in the 18 (50%) patients reevaluated. There was a significant improvement in continence scores and a significantly decreased incidence of UTIs after ELA. The median follow-up time was 34 months (range, 8 to 57 months). CONCLUSIONS AND CLINICAL RELEVENCE: The results of the present study indicated that ELA provided an effective, safe, and minimally invasive treatment option for various VVSRs in dogs, avoiding the need for more invasive surgery.