Histochemical demonstration of hyaluronic acid molecules by alcian blue

Specimens of vitreous humour (monkey eye), Wharton jelly (human umbilical cord) and commercial hyaluronates were immersed in buffered fixative solutions containing either aldehydes and Alcian Blue, or aldehydes and Alcian Blue with MgCl2 as electrolyte. Two MgCl2 concentrations were used, 0.025 M and 0.3 M. Immersion in both solutions induced formation of precipitates which were postfixed in OsO4, dehydrated and embedded for thin section electron microscopy. The use of the same fixative solution produced morphologically comparable precipitates from all three materials. The precipitates, especially after fixation in the presence of electrolyte, were composed of linear, unbranched filaments, frequently aggregated into bundles. The filaments were considered to be molecules of hyaluronic acid.