Between May 1983 and December 1989, 2,921 donor eyes received by the Minnesota Lions Eye Bank underwent a decontamination protocol using 10% Betadine (1% povidone-iodine) solution. Positive pretreatment limbal cultures were obtained on 52.1% of donor eyes. Posttreatment limbal cultures demonstrated a 76.1% reduction in microbial growth, including an 85.7% reduction in Candida species. Streptococcus species were reduced by 76.7% and coagulase-negative Staphylococcus were reduced by 76.1%. In addition, of 250 donor rims obtained at the time of surgery, two had coagulase-negative. Staphylococcus present that were also present after the decontamination procedure. In no cases did endophthalmitis occur during this study. This study demonstrates that this protocol using 10% Betadine solution is extremely effective in decontaminating donor globes of aerobic bacteria and fungi. However, gentamicin-resistant organisms survived this protocol, emphasizing the need to develop storage media containing a broader-spectrum antibiotic agent than gentamicin.