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The Histochemical Journal

, Volume 6, Issue 1, pp 25–34 | Cite as

The role of phosphotungstic and phosphomolybdic acids in connective tissue staining I. Histochemical studies

  • Mona M. Everett
  • William A. Miller
Papers

Synopsis

An investigation of the role of phosphotungstic and phosphomolybdic acids in Mallory-like trichrome methods showed unexpectedly that, rather than acting as mordants to anionic dyes, these polyacids selectively blocked staining of all tissue components other than connective tissue fibres to Aniline Blue and other similar fibrereactive dyes. Connective tissue components were found to contain residues resembling histidine that are easily accessible to anionic dyes. Blocking towards typical anionic dyes for demonstrating plasma proteins, such as Biebrich Scarlet, was also demonstrated but was less complete. The blockade of both types of dye was labile if the staining times were extended; plasma dyes were more sensitive than fibre dyes in this respect. Histochemical reactions for tyrosine residues were blocked. In connective tissue, phosphotungstic acid did not block histidine residues demonstrable by the coupled tetrazonium reaction with previous iodination. Thus it is postulated that differential trichrome staining occurs by binding of Aniline Blue to basic residues in the connective tissue not blocked by phosphotungstic acid and subsequent replacement of the blocking agent by an anionic dye. The binding of phosphotungstic acid to both epithelium and connective tissue was demonstrated by the quenching of autofluorescence in these regions and by the reduction of the bound PTA to blue coloured products with titanium trichloride.

Keywords

Connective Tissue Histidine Trichloride Tissue Component Phosphotungstic Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Adams, C. W. M. (1957). Ap-dimethylaminobenzaldehyde-nitrite method for histochemical demonstration of tryptophan and related compounds.J. clin. Path. 10, 56–62.Google Scholar
  2. Baker, J. R. (1956). The histochemical recognition of phenols, especially tyrosine.Quart. Jl. microsc. Sci. 97, 161–4.Google Scholar
  3. Barrnett, R. J. &Seligman, A. M. (1954). Histochemical demonstration of sulphydryl and disulfide groups of protein.J. natn. Cancer Inst. 14, 769–803.Google Scholar
  4. Brain, E. B. (1962). A new method for preparation of decalcified sections of human enamelin situ.Archs. oral Biol. 7, 757–60.Google Scholar
  5. Bullmer, D. (1962). Observations on histological methods involving the use of phosphotungstic and phosphomolybdic acids, with particular reference to staining with phosphotungstic acid-hematoxylin.Quart. Jl. microsc. Sci. 103, 311–23.Google Scholar
  6. Everett, M. M., Miller, W. A. & Staple, P. H. (1971). Amino acid histochemistry of fetal calf and newborn human enamel matrix.Abstract No. 624.Annual meeting of International Association for Dental Research, Chicago.Google Scholar
  7. Glick, D. &Scott, J. E. (1970). Phosphotungstic Acid not a stain for polysaccharide.J. Histochem. Cytochem. 18, 455.Google Scholar
  8. Goland, P., Scheiman-Tagger, E. &Engel, M. (1965). Enamel preservation during decalcification following fixation by some reactive halogen compounds.J. dent. res. 44, 342–9.Google Scholar
  9. Goland, P. P., Burlakow, P. S. &Grand, N. G. (1967). Cyanuric chloride for improved cytological fixation.Acta cytologica 11, 267–71.Google Scholar
  10. Lillie, R. G. (1965).Histopathologic Technique and Practical Histochemistry, p. 548. New York: McGraw-Hill.Google Scholar
  11. Landing, B. H. &Hall, H. E. (1956). Selective demonstration of histidine.Stain Technol. 31, 197–200.Google Scholar
  12. Miller, W. A. &Everett, M. M. (1972). Amino acid histochemistry of developing and diseased dentine with comparisons to bone.Caries Res. 6, 280 (Abstract).Google Scholar
  13. Palladini, G., Lauro, G. &Basile, A. (1970). Observations sur la spécificité de la coloration aux acides phosphotungstique et phosphomolybdique.Histochemie 24, 315–21.Google Scholar
  14. Pearse, A. G. E. (1968).Histochemistry, Theoretical and Applied, Vol. I, Chapter 6. Boston: Little, Brown.Google Scholar
  15. Pease, D. C. (1966). Polysaccharides associated with the exterior surface of epithelial cells: Kidney, intestine, brain.J. Ultrastruct. Res. 15, 555–88.Google Scholar
  16. Pease, D. C. (1970). Phosphotungstic acid as a specific electron stain for complex carbohydrates.J. Histochem. Cytochem. 18, 455–8.Google Scholar
  17. Puchtler, H. &Isler, H. (1958). The effect of phosphomolybdic acid on the stainability of connective tissue by various dyes.J. Histochem. Cytochem. 6, 265–70.Google Scholar
  18. Quintarelli, G., Zito, R. &Cifonelli, J. A. (1971a). On phosphotungstic acid staining. I.J. Histochem. Cytochem. 19, 641–7.Google Scholar
  19. Quintarelli, G., Cifonelli, J. A. &Zito, R. (1971b). On phosphotungstic acid staining. II.J. Histochem. Cytochem. 19, 648–53.Google Scholar
  20. Scott, J. E. (1971). Phosphotungstate: A ‘universal’ (non-specific) precipitant for polar polymers in acid solution.J. Histochem. Cytochem. 19, 689–95.Google Scholar
  21. Scott, J. E. &Glick, D. (1971). The invalidity of ‘phosphotungstic acid as a specific electron stain for complex carbohydrates’.J. Histochem. Cytochem. 19, 63–4.Google Scholar
  22. Terner, J. Y. (1964). Phosphotungstic acid-hematoxylin; spectrophotometry of the lake in solution and in stained tissue.Stain Technol. 39, 141–53.Google Scholar

Copyright information

© Chapman and Hall Ltd 1974

Authors and Affiliations

  • Mona M. Everett
    • 1
  • William A. Miller
    • 1
  1. 1.Department of Oral BiologyState University of New York at BuffaloSnyderUSA

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