The middle ear muscles and their function have not yet been fully explored. The statement of Lawrence, for example, that the tensor tympani muscle of humans might have a dual innervation has never been proven or disproven. The question is of great interest; in our opinion, it represents one of the key questions in the putative afferent feedback loop of the middle ear muscles in humans. A light microscopic study was performed on 16 tensor tympani muscles taken from 11 cadavers. Six muscles were taken out in toto and stained according to the modified method of Sihler. The remaining 10 muscles were dehydrated and embedded in paraffin. In 5 of these muscles, complete transverse serial sections were made on a microtome at 7 microm and alternately stained by silver impregnation, S-100 protein immunohistochemistry, and ferric oxide. In the remaining 5 muscles, complete longitudinal serial sections were made at 10 microm. These sections were alternately stained by the methods of Cason and Maskar. Neither the surgical microscopic investigation nor the light microscopic investigation revealed any innervation to the human tensor tympani muscle other than the one arising from the mandibular branch of the trigeminal nerve. Our findings, apart from the fact that they clearly refute an unproven hypothesis, might represent another small step toward understanding the innervation of the tensor tympani muscle.