Twenty-four pair of eyes donated to the Minnesota Lions Eye Bank were studied to determine the effect of corneal procurement methods on tissue quality. Eyes studied were ineligible for transplantation because of a preexisting medical condition other than sepsis or age of > years. The procurement technique was randomized for each donor. One cornea was procured in situ (IS), whereas the fellow eye was enucleated and processed in the laboratory (EN). Procurement protocols were standard Eye Bank Association of America methods. Tissue characteristics were scored according to standard eye bank protocols. Cultures were performed at the time of tissue procurement and following storage for 7 days in Dexol media. With the exception of endothelial striae, no statistical difference was found between groups for any tissue characteristics. The average score for endothelial striae in the IS group was greater than twice that of the EN group. Initial cultures were positive in 10 of 24 in the IS group and four of 24 in the EN group. Each group had three positive end-storage cultures. These results demonstrate superior tissue decontamination after initial processing and less endothelial cell trauma with standard enucleation when compared to in situ corneal excisions.