Skip to main content

On the mechanism of sequence iron-hematein stains

Summary

As a prerequisite for the histochemical study of sequence iron-“hematoxylin” stains the iron alum-acidified hematein procedure was developed which does not require differentiation.

Histochemical blocking and extraction procedures demonstrated that carboxyl and hydroxyl groups are essential for the binding of cationic iron.

The iron alum-Prussian blue reaction colored collagen, reticulum fibers and basement membranes more intensely than muscle fibers. Treatment of tissue sections mordanted in iron alum with the acidified hematein solvent resulted in practically complete removal of iron from all tissue structures. It must therefore be concluded that the selective staining of muscle fibers, terminal bars and related structures with sequence iron-hematein stains is not due to high affinity of iron for these tissue components.

Observations by R. and M. Heidenhain on sequence hematoxylin-potassium dichromate and hematoxylin-alum stains and data from modern textile chemistry indicate that the staining patterns obtained with metal-hematein sequence stains are determined by the affinity of the hematein moiety for certain tissue structures.

This is a preview of subscription content, access via your institution.

References

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

    Google Scholar 

  • —, J. N. Desai, D. J. Duff, C. H. Giles, S. K. Jain, and I. R. Macneal: A study of certain natural dyes. I. The adsorption of brazilwood and logwood colouring matters by fibers. J. Soc. Dyes Col. 70, 392–401 (1954 a).

    Google Scholar 

  • Bailey, K.: Structure proteins. II. Muscle. In: The proteins, vol. II, part B (H. Neurath, K. Bailey, ed.). New York: Academic Press 1954.

    Google Scholar 

  • Baker, J. R.: The histochemical recognition of lipine. Quart. J. micr. Sci. 87, 441–470 (1946).

    Google Scholar 

  • —: Principles of biological microtechnique. New York: John Wiley & Sons Inc. 1960.

    Google Scholar 

  • Benda, C.: Über eine neue Färbemethode des Centralnervensystems, und Theoretisches über die Hämatoxylinfärbung. Arch. Anat. Physiol., Phys. Abt., 1886, 562–564.

  • Gaunt, J. F.: Some recent observations on the theory of the dyeing of wool with chrome dyes. J. Soc. Dyers Col. 65, 429–433 (1949).

    Google Scholar 

  • Gerstner, H.: Die Chemie der Applikation von Komplexfarbstoffen. Berlin: Akademie-Verlag 1959.

    Google Scholar 

  • Giles, C. H.: The mechanism of the mordanting of animal fibers. J. Soc. Dyers Col. 60, 303–315 (1944).

    Google Scholar 

  • Gomori, G.: Microscopic Histochemistry. Chicago: Chicago University Press 1952.

    Google Scholar 

  • Gurd, F. R. N., and P. E. Wilcox: Complex formation between metallic cations and proteins, paptides and amino acids. Advanc. Protein Res. 11, 311–427 (1956).

    Google Scholar 

  • Gustavson, K. H.: The chemistry and reactivity of collagen. New York: Academic Press Inc. 1956.

    Google Scholar 

  • Harms, H.: Handbuch der Frabstoffe für die Mikroskopie, Teil II, 2. Liefg. Kamp-Lintfort: Staufen-Verlag 1957.

    Google Scholar 

  • Heidenhain, M.: Beiträge zur Kenntnis der Topographie und Histologie der Kloake und ihrer drüsigen Adnexe bei den einheimischen Tritonen. Arch. mikr. Anat. 35, 173–274 (1890).

    Google Scholar 

  • —: Neue Untersuchungen über die Centralkörper und ihre Beziehungen zum Kern- und Zellenprotoplasma. Arch. mikr. Anat. 43, 423–758 (1894).

    Google Scholar 

  • —: Hämatoxylin-Chromsalze. In: Enzyklopädie der Mikroskopischen Technik, Bd. I. Berlin u. Wien: Urban & Schwarzenberg 1910.

    Google Scholar 

  • Heidenhain, R.: Eine neue Verwendung des Hämatoxylin. Arch. mikr. Anat. 24, 468–470 (1885).

    Google Scholar 

  • Kendrew, J. C.: Structure proteins. I. In: The proteins, vol. II, part B (H. Neurath and K. Bailey, ed.). New York: Academic Press 1954.

    Google Scholar 

  • —, R. E. Dickerson, B. E. Strandberg, R. G. Hart, D. R. Davies, D. C. Phillips, and V. C. Shore: Structure of myoglobin. Nature (Lond.) 185, 422–427 (1960).

    Google Scholar 

  • Klotz, I. M.: Protein interactions. In: The proteins, vol. I, part B (H. Neurath, and K. Bailey, ed.). New York: Academic Press 1953.

    Google Scholar 

  • Landsteiner, K., and J. van der Scheer: Serological differentiation of steric isomers. J. exp. Med. 48, 315–320 (1928).

    Google Scholar 

  • Laskowski, M., and H. A. Scheraga: Thermodynamic considerations of protein reactions. I. Modified reactivity of polar groups. J. Amer. chem. Soc. 76, 6305–6319 (1954).

    Google Scholar 

  • Lillie, R. D.: Histopathologic technic and practical histochemistry. New York: The Blakiston Co., Inc. 1954.

    Google Scholar 

  • Low, B. W.: The structure and configuration of amino acids, peptides and proteins. In: The proteins, vol. I, part A (H. Neurath, and K. Bailey, ed.). New York: Academic Press 1953.

    Google Scholar 

  • Nitschke, G.: Chemische Technologie und Praxis der Färberei, Bd. II. Berlin: VEB Verlag Technik 1958.

    Google Scholar 

  • Otto, G.: Das Färben des Leders. Darmstadt: Eduard Roether 1962.

    Google Scholar 

  • Pearse, A. G. E.: Histochemistry, theoretical and applied. Boston: Little, Brown & Co. 1961.

    Google Scholar 

  • Perutz, M. F., M. G. Rossmann, A. F. Cullis, H. Muirhead, G. Will, and A. C. T. North: Structure of hemoglobin. Nature (Lond.) 185, 416–422 (1960).

    Google Scholar 

  • Puchtler, H., and F. Sweat: Amidoblack as a stain for hemoglobin. Arch. Path. 73, 245–249 (1962).

    Google Scholar 

  • —: Histochemical specifity of staining methods for connective tissue fibers: Resorcinfuchsin and van Gieson's picro-furchsin. Histochemie 4, 24–34, 1964.

    Google Scholar 

  • — and M. Levine: On the binding of Congo red by amyloid. J. Histochem. Cytochem. 10, 355–364 (1962).

    Google Scholar 

  • Race, E., F. M. Rowe, and J. B. Speakman: The nature of the dye-chromium fibre complex in the case of wool dyed with certain chrome mordant dyes. J. Soc. Dyers Col. 62, 372–382 (1946).

    Google Scholar 

  • Rath, H.: Lehrbuch der Textilchemie, 2. Aufl. Berlin-Göttingen-Heidelberg: Springer 1963.

    Google Scholar 

  • Romeis, B.: Mikroskopische Technik. München: Leibniz 1948.

    Google Scholar 

  • Scheraga, H. A.: Protein structure. New York: Academic Press 1961.

    Google Scholar 

  • Schweigger-Scseidel, F.: Über die Grundsubstanz und die Zellen der Hornhaut des Auges. Ber. der Königl. sächs. Ges. d. Wissensch., math.-phys. Kl. 1869. Quoted from G. Schwalbe, Beiträge zur Kenntnis der Drüsen in den Darmwandungen, in's Besondere der Brunner'schen Drüsen. Arch. mikr. Anat. 8, 92–140 (1872).

  • Seki, M.: Zur physikalischen Chemie der histologischen Färbung. IV. Über die Hämateinfärbung. Folia anat. jap. 11, 15–36 (1933).

    Google Scholar 

  • Smythe, C. V., and C. L. A. Schmidt: Studies on the mode of combination of iron with certain proteins, amino acids and related compounds. J. biol. Chem. 88, 241–269 (1930).

    Google Scholar 

  • Springall, H. D.: The structural chemistry of proteins. London: Butterworth Sci. Publ. 1954.

    Google Scholar 

  • Szent-Györgyi, A.: Muscle research. Science 128, 699–702 (1958).

    Google Scholar 

  • Venkataraman, K.: The chemistry of synthetic dyes, vol. 1. New York: Academic Press, Inc. 1952.

    Google Scholar 

  • Vickerstaff, T.: The physical chemistry of dyeing. London: Oliver & Boyd 1954.

    Google Scholar 

  • Wigglesworth, V. B.: The role of iron in histological staining. Quart. J. micr. Sci. 93, 105–118 (1952).

    Google Scholar 

  • Zimmermann, K. W.: Beiträge zur Kenntnis einiger Drüsen und Epithelien. Arch. mikr. Anat. 52, 552–706 (1898).

    Google Scholar 

  • Zipkin, R.: Beiträge zur Kenntnis der gröberen und feineren Strukturverhältnisse des Dünndarms von Inuus Rhesus. Anat. H. 23, 113–185 (1903).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Puchtler, H., Sweat, F. On the mechanism of sequence iron-hematein stains. Histochemie 4, 197–208 (1964). https://doi.org/10.1007/BF00290864

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00290864

Keywords

  • Iron
  • Muscle Fiber
  • Basement Membrane
  • Complete Removal
  • Staining Pattern