Ultrahistochemical study on the ruthenium red surface staining. II. Nature and affinity of the electron dense marker

The electron-dense marker which is thought to produce the ruthenium red surface staining is studied. This stain is prepared under conditions which should give its rise in cell surface membrane, and its nature and charge are tested electrophoretically and by measuring the turbidity, respectively. It is a positive colloid resulting from the recharging of colloidal osmium dioxide by RR polycations. Controls on the affinity are carried out by applying positive sol to gelled agarose sections containing hyaluronic acid, polyvinyl sulfate or polylysine. Controls are also carried out on ascites Ehrlich carcinoma and Zajdela ascites hepatoma cells subjected to prior enzymatic and chemical treatments. It is found that the osmium-RR system visualizes all acidic groups in the outer hydrophilic leaflet, that is the greater part of compounds in this external cell layer. A model is presented for the mechanism underlying its rise in cell surface membrane.