[Investigations on the glycocalyx with toluidin blue staining (author's transl)]

The study was concerned with the molecular structure of the erythrocyte membrane and its anisotropic staining with toluidine blue. Enzymatic or chemical degradation impairs the topo-optical staining. Decreased anisotropy results also from digitonin treatment of glutaraldehyde fixed cells. Incubation in a mixture of chloroform and methanol abolishes the anisotropic staining with toluidine blue, but it has been restored by the aldehyde bisulfite-procedure. The efficacy of a treatment with chloroform-methanol is greatly reduced int eh case of agglutinated erythrocytes. The reversibly collapsed glycocalyx is devoid of topo-optical staining. The findings refer to the spatial orientaiton of glycoproteins at the cell surface. Degradation or loss and rearrangement, respectively, change the glycocalyx structure and likewise deteriorate the condition for the anisotropic staining. Alterations of the lipid layer of the membrane may result in similar effects. There was, however, no indication of an intercalation into the lipid layer of dyestuff molecules. In conclusion, the topo-optical toluidine blue staining is considered a powerful method aiding studies of the glycocalyx of erythrocytes (and presumably of other types of cells too).