[Investigation of the anisotropy of glycocalyx stained with 1.9-dimethyl methylene blue and N,N'-diethylpseudoisocyanine chloride (author's transl)].
In the present study the anisotropic staining of the erythrocyte membrane with 1.9-dimethyl methylene blue and N,N'-diethylpseudoisocyanine chloride was studied and simultaneously compared with the toluidine blue topo-optical staining. The difference between anisotropic toluidine blue and 1.9-dimethyl methylene blue staining, except after KMnO4-oxidation, was only of quantitative nature. On the contrary, striking differences were observed between N,N'-diethylpseudoisocyanine chloride staining, and toluidine blue or 1.9-dimethyl methylene blue staining. Enzymatic and chemical degradation resulted the disappearance of N,N'-diethylpseudoisocyanine chloride staining. Following these treatment membrane birefringence could be restored by aldehyde bisulfate and/or KMnO4-oxidation, while the N,N'-diethylpseudoisocyanine chloride staining was restored only after KMnO4-oxidation. After methylation or acetylation the membrane birefringence disappears, while after KMnO4-oxidation both topo-optical reactions return. The digitonin reaction brought about a rearrangement of the glycocyalyx components. The results draw attention to the spatial orientation of the glycoprotein of the erythrocyte membrane. The role of glycocalyx in the three topo-optical reactions was thus clearly demonstrated.
D001718:Birefringence; D004910:Erythrocyte Membrane / Q000648:ultrastructure*; D004912:Erythrocytes / Q000648:ultrastructure*; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D008751:Methylene Blue; D011804:Quinolines; D013194:Staining and Labeling / Q000379:methods*
D006023:Glycoproteins; D008565:Membrane Proteins; D011804:Quinolines; D008751:Methylene Blue
Untersuchungen über die anisotrope 1:9-Dimethyl-Methylenblau- und N,N'-Diäthylpseudoisocyaninchloridfärbung der Glykokalyx.