PURPOSE: To evaluate the outcomes of intravitreal gas and anti-vascular endothelial growth factor (anti-VEGF), with or without tissue plasminogen activator (tPA), injection for submacular hemorrhage (SMH). METHODS: The authors retrospectively enrolled patients with SMH treated with intravitreal gas and anti-VEGF, with or without tPA (tPA and non-tPA groups, respectively), injection between 2014 and 2021, and data were collected at the preoperative visit and at 1-, 3-, 6-, and 12-month follow-ups. The primary outcome was the final best-corrected visual acuity, and the secondary outcomes were central subfield thickness, degree of blood displacement, and incidence of vitreous hemorrhage. RESULTS: Herein, 38 eyes received intravitreal gas, anti-VEGF, and tPA injections, whereas 89 eyes received intravitreal gas and anti-VEGF injections. At 12 months, no significant intergroup difference was detected (P = 0.94), except the slightly greater central subfield thickness change in the non-tPA group (P = 0.03). Complete SMH displacement occurred in 35 (tPA = 92.1%) and 89 (non-tPA = 89.9%) eyes, with higher vitreous hemorrhage incidence in the non-tPA group (odds ratio 7.03, P = 0.004). CONCLUSION: In pneumatic displacement combined with anti-VEGF therapy in SMH, adjunctive tPA use did not alter the visual and anatomical outcomes, although the vitreous hemorrhage risk in large SMH was reduced.