Bacterial lipopolysaccharide (LPS) is an important agent of induction of ocular pathology following corneal injury or wearing of contaminated contact lenses. The mechanism of LPS uptake through the corneal epithelium is unclear, and the role played by inflammatory cells in this phenomenon has not been previously assessed. Fluorescein isothiocyanate-labeled LPS from Escherichia coli was deposited onto the abraded corneas of New Zealand White rabbits. Epifluorescence microscopy of living excised corneas revealed diffuse LPS staining in the epithelial and stromal layers only in the vicinity of the abrasion. In addition, specific cellular uptake of LPS was suggested by fluorescence staining of cells along the abrasion site. In a second series of experiments, an anti-CD18 polyclonal antibody was used to block infiltration of polymorphonuclear neutrophils (PMN) into the cornea. In these experiments, a diffuse distribution of fluorescent LPS was still observed along the abrasion, but the specific cellular uptake was abolished. The findings indicate that LPS enters the cornea via diffuse penetration at sites of injury and that specific cellular uptake of LPS occurs within the cornea via PMN which have migrated into the damaged tissue.