The anatomy of the extraocular muscles was studied in 10 adult opossums (Didelphis albiventris) of both sexes. Eight extraocular muscles were identified: 4 rectus muscles, 2 oblique muscles, the levator palpebrae superioris and the retractor ocular bulbi. The rectus muscles originate very close one to another between the orbital surfaces of the presphenoid and palatine bones. These muscles diverge on the way to their insertion which occurs at about 2 mm from the limbus. The levator palpebrae superioris originates with the dorsal rectus and is positioned dorsally in relation to it. The retractor ocular bulbi forms a cone which embraces the optic nerve and is located internally in relation to the rectus muscles. The dorsal oblique originates on the presphenoid bone and after a tendinous trajectory through a trochlea on the medial wall of the orbit, inserts into the ocular bulb. The only muscle arising from the anterior orbital floor is the ventral oblique. The main nerve supply for these muscles is the oculomotor, except for the dorsal oblique which is innervated by the trochlear nerve, and the lateral rectus which is innervated by the abducens nerve. The retractor ocular bulbi receives branches from the inferior division of the oculomotor nerve and some branches from the abducens nerve.