Abstract. A reproducible animal model is necessary to examine the use of antimicrobial agents for prophylaxis and treatment of bacterial endophthalmitis. We determined the minimum inoculum size of S. aureus and P. aeruginosa that consistently produced endophthalmitis when injected into aphakic rabbit eyes immediately following surgery. Both anterior chamber and intravitreal injections were examined. For S. aureus, an intravitreal inoculum of 19.3 ± 7.5 CFU and an anterior chamber inoculum of 50.5 ± 4.0 CFU were required. For P. aeruginosa, an intravitreal inoculum of 5.5 ± 2.6 CFU and an anterior chamber inoculum of 97.5 ± 10.7 CFU consistently produced a fulminant infection. Lower inocula of both bacteria produced endophthalmitis in both locations, but the effect was inconsistent.