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The influence of inorganic salts when staining with preformed metal complex dyes

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Summary

The influence of various inorganic salts on the staining of tissue sections by a variety of preformed metal complex dyes (MCDs) has been studied. The staining patterns resulting were found to be extremely complex, though certain generalisations are possible.

Usually the staining of basophilic tissue components (e. g. cartilage matrix and cell nuclei) was reduced as the amount of salt in the dyebath was increased. On the other hand the staining of some acidophilic tissue components (especially elastic fibres) was increased as the salt concentration rose. Both these effects can be rationalised by use of the Donnan equilibrium.

The limited occurrence of salt-induced increases in staining intensity may be attributed in part to the extremely low rates of staining observed with acidophilic substrates.

The decreases and increases in staining seen were to some extent dependant on the particular salt used, and presented an extremely complex picture. One general effect seen was that salts of Al3+, Cr3+, and Mg2+ had a greater inhibitory effect on staining than salts of K+ and Na+. This is explicable in terms of coordinative binding of the polyvalent ions by tissue substrates.

On the basis of the information obtained comments were possible on various topics of biological staining interest. Thus it was concluded that of the MCDs studied only the aluminon-chromium complex showed evidence of binding by a “mordanting” mechanism. Further, it was found that the Critical Electrolyte Concentration theory was commonly inapplicable when staining tissue sections with MCDs.

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References

  • Arshid, F. M., Connelly, R. F., Desai, J. N., Fulton, R. G., Giles, C. H., Kefalas, J. C.: A study of certain natural dyes. II. The structure of the metallic lakes of the brazilwood and logwood colouring matters. J. Soc. Dyers Colour. 70, 402–412 (1954)

    Google Scholar 

  • Baker, J. R.: Principles of biological microtechnique. London: Methuen 1958

    Google Scholar 

  • Baker, J. R.: Experiments on the action of mordants. I. “Singlebath” mordant dyeing. Quart. J. micr. Sci. 101, 255–272 (1960)

    Google Scholar 

  • Berube, G. R., Clark, G.: Characterization and separation of some of the co-ordination compounds of chromium and aluminon. Stain Technol. 39, 329–336 (1964)

    Google Scholar 

  • Berube, G. R., Powers, M. M., Clark, G.: The cationic chelate of chromium and aluminon as a selective nuclear stain. Stain Technol. 39, 337–338 (1964)

    Google Scholar 

  • Berube, G. R., Powers, M. M., Kerkay, J., Clark, G.: The gallocyanin-chrome alum stain, influence of methods of preparation on its activity and separation of active staining compound. Stain Technol. 41, 73–81 (1966)

    Google Scholar 

  • Bobtelsky, M., Ben-Bassat, A.: The aurintricarboxylates of aluminium, iron and chromium. Composition, structure and analytical use. A heterometric study. Anal. chim. acta 14, 344–355 (1956)

    Google Scholar 

  • Coates, E.: Aggregation of dyes in aqueous solutions. J. Soc. Dyers Colour. 85, 355–367 (1969)

    Google Scholar 

  • Culling, C. F. A.: Handbook of histopathological techniques, 2nd ed. London: Butterworths 1963

    Google Scholar 

  • Dutt, M. K.: Celestine blue B as a specific stain for cytochemical detection of nucleic acid. Acta histochem. (Jena) 44, 1–8 (1972)

    Google Scholar 

  • Einarson, L.: A method for progressive staining of Nissl and nuclear substances in nerve cells. Amer. J. Path. 8, 295–305 (1932)

    Google Scholar 

  • Feitelson, J.: Specific effects in the interaction between ion-exchange resins and amino acid cations. Influence of resin cross-linkage. J. Physical Chem. 67, 2544–2547 (1963)

    Google Scholar 

  • Goldstein, D. J.: Correlation of size of dye particle and density of substrate, with special reference to mucin staining. Stain Technol. 37, 79–93 (1962)

    Google Scholar 

  • Goldstein, D. J.: An approach to the thermodynamics of histological dyeing, illustrated by experiments with azure A. Quart. J. micr. Sci. 104, 413–439 (1963)

    Google Scholar 

  • Goldstein, D. J., Horobin, R. W.: Rate factors in staining by alcian blue Histochemical J., 6 (1974, in press).

  • Grenacher, H.: Einige Notizen zur Tinktionstechnik, besonders zur Kernfärbung. Arch. mikr. Anat. 16, 463–471 (1879)

    Google Scholar 

  • Hansen, F. C. C.: Über Eisenhämatein, Chromalaunhämatein, Tonerdealaunhämatein, Hämateinlösungen und einige Cochenillefarblösungen. Z. wiss. Mikr. 22, 45–90 (1905)

    Google Scholar 

  • Hartley, F. R.: The uptake of aluminium by wool. Aust. J. Chem. 21, 1013–1022 (1968a)

    Google Scholar 

  • Hartley, F. R.: Studies in chrome mordanting. II. The binding of chromium III cations to wool. Aust. J. Chem. 21, 2723–2737 (1968b)

    Google Scholar 

  • Heath, I. D.: Observations on a highly specific method for the histochemical detection of sulphated mucopolysaccharides, and its possible mechanisms. Quart. J. micr. Sci. 103, 457–475 (1962)

    Google Scholar 

  • Horobin, R. W.: Analysis and purification of biological stains by gel filtration. Stain Technol. 46, 297–304 (1971)

    Google Scholar 

  • Horobin, R. W., Bennion, P. J.: The interrelation of the size and substantivity of dyes; the role of van der Waals attractions and hydrophobic bonding in biological staining. Histochemie 33, 191–204 (1973)

    Google Scholar 

  • Horobin, R. W., Bennion, P. J., Goldstein, D. J.: Effect of salt in the dyebath on the intensity of staining of tissue sections by acid and basic dyes. Proc. Royal Micro. Soc. 8, 35–36 (1973)

    Google Scholar 

  • Horobin, R. W., Goldstein, D. J.: The influence of salt on the staining of tissue sections with basic dyes: an investigation into the general applicability of the critical electrolyte concentration theory Histochemical J., 6 (1974, in press).

  • Lillie, R. D.: H. J. Conn's biological stains, 8th ed. Baltimore: Williams & Wilkins 1969

    Google Scholar 

  • Lillie, R. D., Bergeaux, G., Henderson, R.: Hot chromation oxyphilia. Histochemie 11, 332–349 (1967)

    Google Scholar 

  • Marshall, P. N., Horobin, R. W.: The chemical nature of the gallocyanin-chrome alum staining complex. Stain Technol. 47, 155–161 (1972)

    Google Scholar 

  • Marshall, P. N., Horobin, R. W.: The mechanism of action of “mordant” dyes — a study using preformed metal complexes. Histochemie 35, 361–371 (1973)

    Google Scholar 

  • Marshall, P. N., Horobin, R. W.: Measurements of the affinities of basic and “mordant” dyes for various tissue substrates. Histochemie (1973b, in press)

  • Meloan, S. N., Valentine, L. S., Puchtler, H.: On the structure of carminic acid and carmine. Histochemie 27, 87–95 (1971)

    Google Scholar 

  • Meybeck, J., Galafassi, P.: The effect of hydrocarbon substituents in azo dyes on wool dyeing. In Applied Polymer Research Symposium No 18. Proceedings of the 4th International Wool Textile Research Conference Pt. 1. New York: Interscience 1971

    Google Scholar 

  • Price, R.: The chemistry of metal complex dyestuffs. In: The Chemistry of synthetic dyes Part III, ed. Venkataraman, K. New York: Academic Press 1970

    Google Scholar 

  • Rattee, I. D.: The use of non-ionic level dyeing assistants with chrome-complex dyes. J. Soc. Dyers Colour. 69, 288–295 (1953)

    Google Scholar 

  • Ringertz, N. R.: Cytochemical properties of nuclear proteins and deoxyribonucleoprotein complexes in relation to nuclear function. In Handbook of molecular cytology, ed. Limade-Faria, A. London: North Holland 1969

    Google Scholar 

  • Sandritter, W., Kiefer, G., Rick, W.: Gallocyanin-chrome alum. In: Introduction to quantitative cytochemistry, ed. Wied, G. L. New York: Academic Press 1966

    Google Scholar 

  • Scott, J. E.: Patterns of specificity in the interaction of organic cations with acid mucopoly-saccharides. In: The chemical physiology of mucopolysaccharides, ed. Quintarelli, G. London: Churchill 1968

    Google Scholar 

  • Sillén, L. G., Martell, A. E.: Stability constants. Special Publications Nos. 17 and 25. London: The Chemical Society 1964 and 1971

    Google Scholar 

  • Sorvari, T. E.: Binding of ferric ions to nuclei and other tissue sites in sections stained for sulphated mucosubstances by the high-iron diamine method. Stain Technol. 47, 245–248 (1972a)

    Google Scholar 

  • Sorvari, T. E.: Histochemical observations on the role of ferric chloride in the high-iron diamine technique for localizing sulphated mucosubstances. Histochem. J. 4, 193–204 (1972b)

    Google Scholar 

  • Thompson, S. W.: Selected histochemical and histopathological methods. Springfield: Thomas 1966

    Google Scholar 

  • Tolgyesi, W. S., Cottington, E. M.: The binding of aluminium by hair. In Applied Polymer Symposium No. 18. Proceedings of the 4th International Wool Textile Research Conference Pt. 1. New York: Interscience 1971

    Google Scholar 

  • Whewell, C. S., Stevens, C. B., Amin, S. C.: Observations on the dyeing behaviour of chemically modified wools. In Applied Polymer Research Symposium No. 18. Proceedings of the 4th International Wool Textile Research Conference Pt. 1. New York: Interscience 1971

    Google Scholar 

  • Wolstenholme, G. E. W., O'Connor, M.: CIBA Symposium on the chemistry and biology of mucopolysaccharides. London: Churchill 1958

    Google Scholar 

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  1. P. N. Marshall

    Present address: Department of Haematology, Royal Postgraduate Medical School, Hammersmith Hospital, W12, London

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  1. Department of Human Biology and Anatomy, The University, Sheffield, UK

    P. N. Marshall & R. W. Horobin

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  2. R. W. Horobin
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Marshall, P.N., Horobin, R.W. The influence of inorganic salts when staining with preformed metal complex dyes. Histochemie 37, 299–311 (1973). https://doi.org/10.1007/BF00274966

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