, Volume 43, Issue 4, pp 333–348 | Cite as

Aldehyde bisulfite-toluidine blue (ABT) staining as a topo-optical reaction for demonstration of linear order of vicinal OH groups in biological structures

  • Gy. Romhányi
  • Gy. Deák
  • J. Fischer


The aldehyde-bisulfite-toluidine blue reaction followed by poststaining stabilization with potassium ferricyanide (ABT) is described as a topo-optical, oriented staining reaction of the vicinal OH groups of complex carbohydrates in biological structures such as polysaccharides, glycoproteins and glycolipids.

The birefringence as induced by the oriented dye binding as a result of ABT is indicative of linear order of the vicinal OH groups and, in turn, provides information on the ultrastructural pattern of carbohydrate moieties in biological substances, which pattern is often not demonstrable by other ultrastructural methods.

The possibilities of this new approach to the ultrastructural analysis of complex carbohydrates with ABT in a great number of biological substances is demonstrated and its practical value in histopathology discussed.


Public Health Potassium Carbohydrate Aldehyde Polysaccharide 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ainsworth, S. K., Ito, S., Karnovsky, M. J.: Alkaline bismuth reagent for high resolution ultrastructural demonstration of periodate reactive sites. J. Histochem. Cytochem. 20, 995–1005 (1972)Google Scholar
  2. Barka, T., Anderson, P. J.: Histochemistry. Theory, practice and bibliography, p. 68–95, and p. 139. New York-Evanston-London: Hoeber Med. Div. Harper & Row, Publ. Inc. 1963Google Scholar
  3. Bradbury, S., Stoward, P. J.: The specific cytochemical demonstration in the electron microscope of periodate-reactive mucosubstances and polysaccharides containing vicglycol groups. Histochemie 11, 71–80 (1967)Google Scholar
  4. Csuka, O., Sugár, J.: Electron microscopic studies on the specificity of the PA-silver reaction. Acta Morph. Acad. Sci. Hung. 19, 233–240 (1971)Google Scholar
  5. Deák, Gy., Karlinger, K.: Submicroscopic structure of the corpora amylacea of the central nervous system. Acta morph. Acad. Sci. hung. 20, 77–84 (1972)Google Scholar
  6. DeMarsh, Q. B., Kautz, J.: The submicroscopic morphology of Gaucher cells. Blood 12, 324–335 (1957)Google Scholar
  7. Drochmans, P.: Morphologie du glycogéne. Etude au microscope électronique de colorations négatives du glycogéne particulaire. J. Ultrastruct. Res. 6, 141–163 (1962)Google Scholar
  8. Frey-Wyssling, A.: The fine structure of cellulose microfibrils. Science 119, 80–82 (1954)Google Scholar
  9. Hagen, A.: Pflanzliche Zellwand. In: Molekulare Biologie der Zelle, Hrsg. Bielka, H., S. 293–310. Jena: VEB G. Fischer Verlag 1969Google Scholar
  10. Hall, B. V.: Renal glomerular basement membrane as a macromolecular system forming a complex multiple random-slit molecular filter. Anat. Rec. 151, 356–357 (1965)Google Scholar
  11. Hotchkiss, R. D.: Microchemical reaction resulting in the staining of polysaccharide structures in fixed tissue preparations. Arch. Biochem. 16, 131–141 (1948)Google Scholar
  12. Kefalides, N. A.: Isolation of a collagen from basement membranes containing three identical α-chains. Biochem. biophys. Res. Commun. 45, 226–234 (1971)Google Scholar
  13. Lillie, R. D.: Reticulum staining with Schiff-reagent after oxidation by acidified sodium perjodat. J. Lab. clin. Med. 22, 910–912 (1947)Google Scholar
  14. Luzardo-Baptista, M.: Correlation between molecular structure and glycogen ultrastructure Ann. Histochim. 17, 141–144 (1972)Google Scholar
  15. Malinin, G.: Metachromatic staining of sodium bisulfite addition derivatives of glycogen. J. Histochem. Cytochem. 18, 834–841 (1970)Google Scholar
  16. McManus, J. F. A.: The histological demonstration of mucin after periodic acid. Nature (Lond.) 158, 202 (1946)Google Scholar
  17. Missmahl, H. P.: Doppelbrechung der retikulären Faser und sich hieraus ergebender Nachweis von gerichtet eingelagerten Lipoiden in die retikuläre Faser. Z. Zellforsch. 45, 612 (1957)Google Scholar
  18. Missmahl, H. P., Kübler, W.: Polarisationsoptische Untersuchungen an Gaucherzellen. Frankfurt. Z. Path. 71, 190–193 (1961)Google Scholar
  19. Módis, L.: Topo-optical investigations of mucopolysaccharides. (acid glycosaminoglycans). Polysaccharides. II. In: Handbuch der Histochemie (Graumann u. Neumann, Hrsg.) S. 11–15. Stuttgart: Gustav Fischer Verlag 1974Google Scholar
  20. Movat, H. Z.: Silver impregnation methods for electron microscopy. Amer. J. clin. Path. 35, 528–537 (1961)Google Scholar
  21. Mowry, R. W.: The special value of methods that color both acidic and vicinal hydroxyl groups in the histochemical study of mucins. With revised directions for the colloidal iron stain, the use of alcian blue G8X and their combinations with the periodic acid-Schiff reaction. Ann. N.Y. Acad. Sci. 106, 402–423 (1963)Google Scholar
  22. Musy, J. P., Sprumont, P., Módis, L., de Blasi, V.: Ultrastructure du mucus (cellules calciformes du colon) et des granulations des mastocytes du colon. Histochemie 30, 40–59 (1972)Google Scholar
  23. Rambourg, A.: An improved silver methenamine technique for the detection of periodic acid-reaction complex carbohydrates with the electron microscope. J. Histochem. Cytochem. 15, 409–412 (1967)Google Scholar
  24. Romhányi, Gy.: Über die submikroskopische strukturelle Grundlage der metachromatischen Reaktion. Acta histochem. (Jena) 15, 201–233 (1963)Google Scholar
  25. Romhányi, Gy., Bukovinszky, A., Deák, Gy.: Sulfation as a collagen-specific reaction. The ultrastructure of sulfate collagen, basement membranes and reticulin fibers as shown by topo-optical reactions. Histochemie 36, 123–138 (1973)Google Scholar
  26. Romhányi, Gy., Bukovinszky, A., Deák, Gy.: Selective proteolytic sensitivity of sulfatecollagen and basement membranes. Histochemistry 42, 199–209 (1974b)Google Scholar
  27. Romhányi, Gy., Deák, Gy.: On the ultrastructural organization of biological membranes as shown by topooptical staining reactions. Acta morph. Acad. Sci. hung. 17, 245–272 (1969)Google Scholar
  28. Romhányi, Gy., Deák, Gy.: Topo-optical studies on age pigment, corpora amylacea and senile amyloid-like substances of brain. VII. Internat. Congr. of Neuropath., Budapest, 1974Google Scholar
  29. Romhányi, Gy., Molnár, L.: Optical polarization indicates linear arrangement of rhodopsin oligosaccharide chain in rod disc membranes of frog retina. Nature (Lond.) 249, 486–488 (1974)Google Scholar
  30. Romhányi, Gy., Molnár, L., Németh, A.: Ultrastructural differences in cell membranes of erythrocytes myeloid and lymphoid cells as shown by topo-optical reactions. Histochemistry 39, 261–276 (1974a)Google Scholar
  31. Scheuner, G., Hutschenreiter, J.: Polarisationsmikroskopie in der Histophysik, S. 126–130. Leipzig: G. Thieme 1972Google Scholar
  32. Schmidt, W. J.: Der Wandel der optischen Anisotropie bei topochemischen Reaktionen histologischer Strukturen. Ber. oberhess. Ges. Natur.- u. Heilkunde, Naturwiss. Abt. 23, 56–85 (1947)Google Scholar
  33. Shackleford, J. M.: Histochemical comparison of mucous secretions in Rodent, Carnivore, Ungulate, and Primate major salivary glands. Ann. N.Y. Acad. Sci. 106, 572–582 (1963)Google Scholar
  34. Spiro, R. G.: The structure of the disaccharide unit of the renal glomerular basement membrane. J. biol. Chem. 242, 4813–4823 (1967)Google Scholar
  35. Spiro, R. G., Lucas, F., Rudall, K. M.: Glycosylation of hydroxylysine in collagen. Nature (Lond.) New Biol. 231, 54–55 (1971)Google Scholar
  36. Thiéry, J. P.: Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J. Microscopie 6, 987–1018 (1967)Google Scholar
  37. Thornell, L.-E.: The fine structure of Purkinje fiber glycogen. A comparative study of negatively stained and cytochemically stained particles. J. Ultrastruct. Res. 49, 157–166 (1974)Google Scholar
  38. Vidal, B. C. de, Mello, S.: Macromolecular conformation of the colon mucus as revealed by detection of anisotropic phenomena. Ann. Histochem. 19, 151–156 (1974)Google Scholar
  39. Westphal, O., Lüderitz, O.: Die biologische Bedeutung der chemischen Feinstruktur bakterieller Zellgrenzflächen. Naturwissenschaften 50, 413–426 (1963)Google Scholar
  40. White, J. C., Elmes, P. C., Walsh, A.: Fibrous proteins of pathological bronchial secretions studied by optical and electron microscopy. Deoxyribonucleoprotein and mucoprotein in bronchial secretions. J. Path. Bact. 67, 105–108 (1954)Google Scholar

Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • Gy. Romhányi
    • 1
  • Gy. Deák
    • 1
  • J. Fischer
    • 1
  1. 1.Institute of Pathological AnatomyUniversity Medical SchoolPécsHungary

Personalised recommendations