The use of topical antibiotics as prophylaxis after intravitreal injections is a contentious issue, especially now with increasing use of intravitreal anti-VEGF agents. In the Wills Eye Institute, prior to May 2011, all patients undergoing intravitreal injections in the office setting routinely received post-injection topical antibiotics four times daily for four days. However, the practice of post-injection antibiotics was discontinued practice-wide from September 2011 onwards. Apart from the cessation of post-injection antibiotics, there were no other changes to the technique of preparing for and administering the injections (with a 30- or 31-gauge needle). Topical 5% povidone iodine was applied pre-injection, but sterile draping and eyelash preparation were not used. The authors retrospectively compared the incidence of endophthalmitis following intravitreal injections during and after the post-injection topical antibiotic period. During the 28-month post-injection topical antibiotic period studied, 57,654 injections were administered with 28 cases of suspected endophthalmitis (0.049%) and 10 culture-positive endophthalmitis (0.017%). During the nine month period when post-injection topical antibiotics were discontinued, 34,900 injections were administered with 11 cases of suspected endophthalmitis (0.032%) and four culture-positive endophthalmitis (0.011%). The difference in endophthalmitis rates between the two groups was not statistically significant. This is the largest reported series of intravitreal injections so far, and the results are very compelling in demonstrating that post-injection topical antibiotics do not reduce the risk of endophthalmitis and in fact may even increase the risk of endophthalmitis. The evidence is clear that there is no role for routine post-injection antibiotic prophylaxis.