Purpose <p>There is no established method for visualizing the three-dimensional (3D) structure of the aqueous humor outflow tract. This study attempted to visualize the 3D structures of porcine and human ocular tissues, particularly the aqueous humor outflow tract using a transparency reagent composed of 2, 2-thiodiethanol.</p> Study Design <p>Clinical and experimental.</p> Methods <p>The porcine eyes were collected in Japan, and the human eyes were imported from the United States. The human eyes were obtained from a 64-year-old Caucasian woman, arriving 7 days after her death. The specimens were formalin-fixed upon arrival, fluorescently labeled, optically cleared using a transparency-enhancing reagent, and visualized using a confocal microscope.</p> Results <p>Both porcine and human eyes were visualized to the extent that the choroidal vessels were observed on gross examination. The aqueous humor outflow tract was clearly observed as a luminal structure in the porcine eye, mainly depicted by autofluorescence, and in the human eyes as a luminal structure continuing from the trabecular meshwork without fluorescence.</p> Conclusion <p>Observations using transparency-enhancing technology enabled us to obtain 3D images useful for visualizing ocular tissues, especially the aqueous humor outflow tract.</p>