We evaluated the effect of saline irrigation on 38 pairs of donor eyes and determined the relative efficacy of antibiotic rinse/storage (neomycin-polymyxin B-gramicidin) and povidone-iodine immersion. Microbial growth was found in 49 (64.5%) of 76 eyes from which cultures were taken before irrigation, and only four (8.2%) became culture negative after irrigation. Of 27 eyes culture negative before irrigation, 15 (55.6%) became positive after irrigation. One eye of each pair was then assigned randomly to antibiotic rinse/storage and the other eye to 3-minute immersion in 1% povidone-iodine. Both antimicrobial treatments decreased the numbers of positive cultures, with povidone-iodine showing an advantage that did not reach significance. These results demonstrate that saline irrigation of donor eyes in situ does not reduce surface contamination and that povidone-iodine immersion should be considered as an alternative for donor eye decontamination.