The purpose of this study was to compare Optisol to moist chamber storage for maintaining human corneal endothelial barrier function. Human corneas preserved in Optisol were stored for up to 35 days at 4 C. Endothelial carboxyfluorescein permeability (P(ac)) was measured and endothelial ultrastructure was evaluated by electron microscopy. Endothelial P(ac) (x 10(-4) cm/min) of Optisol-stored corneas was 1.7, 2.0, and 3.1 at five, seven, and 14 days, respectively. The P(ac) increased to 6.5 at 35 days of storage. Endothelial P(ac) in moist chamber stored-eyes was 2.6 at two days, and increased to 13.5 14 days of storage. Multiple regressional analysis showed that storage time and donor age affected P(ac); but time from death to enucleation, time from enucleation to storage, or endothelial cell number did not. Electron microscopy showed that endothelial junctions were maintained through two weeks by Optisol. Large areas of cellular destruction were seen after five days of moist chamber storage. These results show that Optisol can preserve endothelial barrier function through 14 days; barrier function is lost by three days of moist chamber storage.