The identification, distribution, and localization of matrix proteins and the proteins associated with normal and degenerated elastic fibers and collagen fibrils of myxomatous chordae tendineae were studied with immunoelectron microscopy. Ultrathin sections of L R White-embedded tissue were processed by indirect immunogold cytochemistry using primary antibodies against human alpha elastin, collagen types I and III, fibronectin, and vitronectin. In normal chordae tendineae, alpha elastin antibody heavily labeled the elastic fibers in spongiosa and fibrosa, but microfibrils around them were not labeled. Antibodies to collagen type I, collagen type III, and fibronectin all labeled the collagen fibers and microfibrils in the spongiosa. Fibronectin antibody labeling was higher than collagen type III, whereas labeling by anticollagen type I was lower. Intense labeling by vitronectin was observed on the microfibrils in the spongiosa and on electron-dense material around elastic fibers in the spongiosa and fibrosa. In myxomatous chordae tendineae, alpha elastin antibody heavily labeled degenerated elastic fibers, previously unidentified reticulated structures, and other moderately electron-dense material, both in the spongiosa and in the fibrosa, but not the electron-dense fibrous material around them. Antibodies to collagen types I, III, and fibronectin heavy labeled electron-dense aggregates of fibrous material. Vitronectin labeling was observed on electron-dense longitudinally running microfibrils and on the electron-dense microfibrils around degenerated elastic fibers.