To determine practice preference of prophylaxis against post-cataract surgery endophthalmitis in Singapore. Prospective nationwide survey of all registered ophthalmologists through telephone, e-mail or postal questionnaires. A response rate of 76.6% was obtained from 121 out of 158 eligible ophthalmologists. Awareness of the recommendations made by the European Society of Cataract and refractive surgeons (ESCRS) endophthalmitis study was noticed in 98 (81.0%) respondents. Out of the 121 respondents, 85 (70.2%) do not use intracameral antibiotic prophylaxis. Of the 36 respondents using intracameral antibiotics, 34 (94.4%) used cefazolin. The reasons cited for not adopting intracameral antibiotic prophylaxis included financial costs, the relatively low risk of endophthalmitis after cataract surgery, the burden of dilution and the fear of toxic anterior segment syndrome. More than half (65, 53.7%) of respondents would use intracameral antibiotic prophylaxis if it were available commercially. The most commonly practiced prophylaxis (94, 77.7%) was preoperative topical antibiotics. The majority of ophthalmologists in Singapore have not adopted the use of intracameral antibiotics, with most preferring the use of pre- and postoperative prophylactic topical antibiotics, despite knowledge of the ESCRS endophthalmitis study.