Anteroposterior and tangential traction on the central retina is an important factor in the pathogenesis of idiopathic macular hole formation. Histological studies have shown that macular holes of different stages can be associated with epiretinal membranes. Such membranes can be removed during surgery for macular holes. We investigated such tissue samples of 11 patients with macular holes in stages II-IV. Light microscopically, the tissue consisted of a thin collagen layer mostly covered by a thin layer of cells. Ultrastructural analysis revealed glial cells and macrophages as cellular components. The collagen can be ascribed to vitreous, inner limiting membrane and newly formed collagen. According to the morphological findings a multilayered tissue structure can be assumed. Macrophages were found on the retinal side of the inner limiting membrane and at the vitreal side of the tissue. Therefore, the macrophages probably originate from the retina as well as from the vitreous as so-called resident hyalocytes. Glial cells covered the inner limiting membrane forming pericellular collagen to which outer vitreous collagen fibrils can be attached. The multilayered membrane structure might possibly be the cause for only partial laminar surgical extraction so that contractile or potentially proliferative tissue residues might be one of the reasons for surgical failures after incomplete membrane peeling.