To evaluate the use of tenecteplase in transoperative phacoemulsification in healthy rabbits, the study was carried out with fifteen New Zealand rabbits, divided into three groups: control group (CG), untreated group (UG) and treated group (TG). UG and TG were operated by phacoemulsification and TG received 50 µg / 0.3 mL of intracameral tenecteplase. The postoperative evaluations were 24 h, 72 h, 7 days, 15 days and 21 days. In TP21 the animals were submitted to euthanasia and aqueous humor samples were collected. No significant differences were observed in the clinical evaluations between CG and TG in relation to incidence rates of intraocular pressure (IOP), corneal edema, fibrin deposits, hyphema, aqueous flare and synechia. In the physicochemical evaluation of the aqueous humor, there were no significant differences between the three groups in relation to pH values and concentrations of chloride ions. The aqueous humor density values were statistically different between CG and the other groups. In the histological evaluation, there were no significant differences between the groups. The use of tenecteplase in transoperative phacoemulsification in rabbits did not present significant differences in terms of clinical, physicochemical and histological parameters.