A 29-year-old woman presented with clinical signs and symptoms of Acanthamoeba keratitis. Scanning slit confocal microscopy revealed a 26-mu-diameter object, resembling an Acanthamoeba cyst, in the anterior stroma. Numerous ovoid objects (possibly inflammatory cells, trophozoites, or altered keratocytes) were present. Normal keratocyte nuclei and the anterior corneal mosaic, readily imaged by scanning slit confocal microscopy of the normal cornea, were noticeably absent. Subsequent corneal biopsy confirmed the diagnosis of Acanthamoeba keratitis.