The use of immunocytochemical techniques in studying the ultrastructure of elastin in vascular smooth muscle cell cultures: I. Anti-alpha-elastin staining pattern

Abstract

We have used cell cultures to localize elastin in the extracellular matrix (ECM) by ultrastructural immunocytochemistry. An ideal model for such studies are neonatal rat aortic smooth muscle cell (SMC) cultures described by Oakes et al. As much as fifty percent of the total protein in the ECM of these cultures is elastin. During recent years, an alternative and most useful application of the antibody staining “sandwich” technique has evolved to indirectly localize tissue antigens on ultrathin sections. The sandwich technique involves the use of a marker employing colloidal gold-labelled IgG intermediate layer acting to detect the presence of primary antibody. In this case, the primary antibody (anti-alpha-elastin) attaches to the epitope(s) of the elastin molecule present on the cut surface of the sectioned elastic fiber. The value of the immunogold (IMG) antibody staining technique lies in its extraordinary ability for detecting the 10-40 nm diameter colloidal gold particle.