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Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions

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Summary

The application of the “critical electrolyte concentration” (CEC) concept to the differentiation of acidic glycosaminglycans (mucopolysaccharides) is described. Alcian Blue 8GX stains with increasing selectivity as increasing amounts of magnesium chloride are incorporated into the dye solution. Model experiments with pure polyanions, or artifically carboxylated, phosphorylated and sulphated liver sections, showed that binding of dye to carboxylate or phosphate groups ceased at low electrolyte concentrations (< 0.3M) whereas dye continued to be held by sulphate ester groups at concentrations five to ten times as high. The similarity to the well established cetylpyridinium system for polyanion fractionation is discussed.

Sections of tissues chosen to contain predominantly or characteristically carboxylated mucins, and/or sulphate ester polyanions showed a staining pattern entirely similar to the model sections. Goblet cell mucin in rat ileum stained at < 0.4M MgCl2, Cartilage at < 0.6M MgCl2, mast cells at < 0.75M, and corneal stroma at < 1.0M. These results are in agreement with the known contents of sialo-mucin, chondroitin sulphate, heparin and keratansulphate, respectively. The conditions in which this principle can be used in a practical technique are described.

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References

  • Antonopoulos, C. A., B. Borelius, S. Gardell, B. Hamnström, and J. E. Scott: The Precipitation of Polyanions by Long-chain Aliphatic Ammonium Compounds. Pt. IV. Elution in Salt Solutions of Mucopolysaccharide-Quaternary Ammonium Complexes adsorbed on a Support. Biochim. biophys. Acta (Amst.) 54, 213–226 (1961).

    Google Scholar 

  • Booij, H. L.: Colloid Chemical Aspects of Metachromasia. Acta histochem. (Jena), Suppl. 1, 37–55 (1958).

    Google Scholar 

  • Heath, I. D.: Staining of Sulphated Mucopolysaccharides. Nature (Lond.) 191, 1370–1371 (1961).

    Google Scholar 

  • Jeanloz, R. W.: The Nomenclature of Mucopolysaccharides. Artrh. and Rheum. 3, 233–237 (1960).

    Google Scholar 

  • Kelly, J. W., G. D. Bloom, and J. E. Scott: Quaternary Ammonium Compounds in Connective Tissue Histochemistry. I. Selective Unblocking. J. Histochem. Cytochem. 11, 791–798 (1963).

    Google Scholar 

  • Landing, H. B., and H. B. Hall: Phosphorylation as a Histochemical Procedure. J. Histochem. Cytochem. 4, 382–387 (1956).

    Google Scholar 

  • Landsmeer, J. M. F.: Some colloid Chemical Aspects of Metachromasia. Influence of pH and Salts on Metachromatic Phenomena evoked by Toluidine Blue in Animal Tissue. Acta physiol. pharmacol. neerl. 2, 112–128 (1951).

    Google Scholar 

  • Laurent, T. C., and A. Anseth: Studies on Corneal Polysaccharides. II. Characterisation. Exp. Eye Res. 1, 99–105 (1961).

    Google Scholar 

  • —, and J. E. Scott: Molecular Weight Fractionation of Polyanions by Cetylpyridinium Chloride in Salt Solutions. Nature (Lond.) 202, 661–662 (1964).

    Google Scholar 

  • Mowry, R. W.: Observations on the Use of Sulphuric Acid in Ether for the Sulphation of Hydroxyl Groups in Tissue Sections. J. Histochem. Cytochem. (Abstract) 6, 82 (1958).

    Google Scholar 

  • Quintarelli, G., J. E. Scott, and M. C. Dellovo: Chemical and Histochemical Properties of Alcian Blue, III. Chemical Blocking and Unblocking. Histochemie 4, 99–112 (1964).

    Google Scholar 

  • Sainte-Marie, G.: A Paraffin Embedding Technique for Studies employing Immunofluorescence. J. Histochem. Cytochem. 10, 250–254 (1962).

    Google Scholar 

  • Saunders, A. M.: Histochemical Identification of Acid Mucopolysaccharides with Acridine Orange. J. Histochem. Cytochem. 12, 164–170 (1964).

    Google Scholar 

  • Scott, J. E.: Aliphatic Ammonium Salts in the Assay of Acidic Polysaccharides from Tissues. Methods of Biochemical Analysis (ed. D. Glick), vol. 8, p. 145–197. New York: Interscience Publ. 1960.

    Google Scholar 

  • - The Fractionation of Polyanions by Long-chain Aliphatic Ammonium Salts. Biochem. J. 78, 24 P (1961).

  • —: The Precipitation of Polyanions by Long-chain Aliphatic Ammonium Salts, part VI. The Affinity of Substituted Ammon Cations for the Anionic Groups of some Biological Polymers. Biochem. J. 84, 270–275 (1962).

    Google Scholar 

  • Scott, J. E., J. Dorling, and G. Quintarelli: Differential Staining of Acid Glycosaminoglycans by Alcian Blue in Salt Solutions. Biochem. J. 90, 4–5 P (1964).

    Google Scholar 

  • Shackleford, J. M.: Carboxylation of Tissue Sections with the Use of p-Hydrazinbenzoio Acid and Succinic Anhydride. J. Histochem. Cytochem. 10, 469–472 (1962).

    Google Scholar 

  • Szirmai, J. A.: Quantitative Approaches in the histochemistry of Mucopolysaccharides. J. Histochem. Cytochem. 11, 24–34 (1963).

    Google Scholar 

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The new and more precise terminology of Jeanloz (1960) is used in preference to the older nomenclature.

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Scott, J.E., Dorling, J. Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions. Histochemie 5, 221–233 (1965). https://doi.org/10.1007/BF00306130

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  • DOI: https://doi.org/10.1007/BF00306130

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