We present eight cases ofAcanthamoebakeratitis. In each case, theAcanthamoebaorganisms were visualized in the epithelium and anterior stroma using tandem scanning confocal microscopy. The organisms were highly reflective, ovoid, and were 10–25 μm in diameter. TheAcanthamoebaorganisms in the human corneas were identical in size and shape toAcanthamoebaorganisms on an agar plate visualized with the same confocal microscope. Confocal microscopy is a useful method for identifying.Acanthamoebaorganisms in vivo within the corneal epithelium and anterior stroma.