A sensitive post-DAB enhancement technique for demonstration of iron in the central nervous system (original) (raw)

Abstract

A technique is described for enhancing the reaction product of the staining reaction for iron in paraffin-embedded tissue from central nervous system (CNS). After amplification of the Prussian Blue staining reaction with 3,3′-diaminobenzidine (DAB), the reaction product was further intensified using a stepwise treatment with silver methenamine, gold chloride and uranyl nitrate (post-DAB treatment). Following the Prussian Blue-DAB staining reaction, iron was seen only in glial cells and choroid plexus epithelial cells, whereas the post-DAB treatment revealed that neurons and endothelial cells of the brain capillaries were also positively stained. The post-DAB treatment resulted additionally in an increased intensity of the reaction product within choroid plexus epithelial cells compared to that obtained in sections subjected only to the Prussian Blue-DAB reaction. The reliability of the method was evaluated using liver sections as positive controls. Furthermore the higher sensitivity of the method was assessed using nitrocellulose filters containing serially diluted iron-saturated transferrin. The post-DAB method is simple and can easily be applied to formalin- or glutaraldehyde fixed, paraffin-embedded nervous and non-nervous tissue.

Access this article

Log in via an institution

Subscribe and save

• Get 10 units per month
• Download Article/Chapter or eBook
• 1 Unit = 1 Article or 1 Chapter
• Cancel anytime Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

• Bloch B, Popovici T, Chouham S, Levin MJ, Tuil D, Kahn A (1987) Transferrin gene expression in choroid plexus of the adult rat brain. Brain Res Bull 18:573–576
  Google Scholar
• Connor JR, Menzies SL (1990) Altered cellular distribution of iron in the central nervous system of myelin deficient rats. Neuroscience 34:265–271
  Google Scholar
• Connor JR, Menzies SL, St. Martin SM, Mufson EJ (1990) Cellular distribution of transferrin, ferritin, and iron in normal and aged human brains. J Neurosci Res 27:595–611
  Google Scholar
• Crowe A, Morgan EH (1992) Iron and transferrin uptake by brain and cerebrospinal fluid in the rat. Brain Res 592:8–16
  Google Scholar
• Dwork AJ, Schon EA, Herbert J (1988) Nonidentical distribution of transferrin and ferric iron in human brain. Neuroscience 27:333–345
  Google Scholar
• Francois C, Nguyen-Legros J, Percheron G (1981) Topographical and cytological localization of iron in rat and monkey brains. Brain Res 215:317–322
  Google Scholar
• Gallayas F, Görcs T, Merchenthaler I (1982) High-grade intensification of the end-product of the diaminobenzidine reaction for peroxidase histochemistry. J Histochem Cytochem 30:183–184
  Google Scholar
• Giometto B, Bozza F, Argentiero V, Gallo P, Pagni S, Piccinno MG, Tavolato B (1990) Transferrin receptors in rat central nervous system — an immunocytochemical study. J Neurol Sci 98:81–90
  Google Scholar
• Goto S, Nagahiro S, Ushio Y, Hofer W (1992) A simple enhancement method for the silver-gold-intensified diaminobenzidine reaction in the light microscopic immunoperoxidase technique. J Histochem Cytochem 40:1423–1425
  Google Scholar
• Hill JM, Switzer RC (1984) The regional distribution and cellular localization of iron in the rat brain. Neuroscience 11:595–603
  Google Scholar
• Jeffries WA, Brandon MR, Hunt SV, Williams AF, Gatter KC, Mason DY (1984) Transferrin receptor on endothelium of brain capillaries. Nature 312:161–163
  Google Scholar
• Møllgård K, Dziegielewska KM, Saunders NR, Zakut H, Soreq H (1988) Synthesis and localization of plasma proteins in the developing human brain. Integrity of the fetal blood-brain barrier to endogenous proteins of hepatic origin. Dev Biol 128:207–221
  Google Scholar
• Nguyen-Legros J, Bizot J, Bolesse M, Pulicani J-P (1980) “Noir de diaminobenzidine”: une nouvelle méthode histochimique de révélation du fer exogéne. Histochemistry 66:239–244
  Google Scholar
• Perl DP, Good PF (1992) Comparative techniques for determining cellular iron distribution in brain tissues. Ann Neurol 32 [Suppl]:S76-S81
  Google Scholar
• Perls M (1867) Nachweis von Eisenoxyd in gewissen Pigmenten. Virchows Arch 39:42–48
  Google Scholar
• Rodríguez EM, Yulis R, Peruzzo B, Alvial G, Andrade R (1984) Standardization of various applications of methacrylate embedding and silver methenamine for light and electron microscopic immunocytochemistry. Histochemistry 81:253–263
  Google Scholar
• Sciot R, Verhoeven G, Van Eyken P, Cailleau J, Desmet VJ (1990) Transferrin receptor expression in rat liver: immunohistochemical and biochemical analysis of the effect of age and iron storage. Hepatology 11:416–427
  Google Scholar
• Scopsi L, Larsson L-I (1986) Increased sensitivity in peroxidase immunocytochemistry. A comparative study of a number of peroxidase visualization methods employing a model system. Histochemistry 84:221–230
  Google Scholar

Download references

Author information

Authors and Affiliations

1. Department of Medical Anatomy and Neuroscience Centre, The Panum Institute, University of Copenhagen, Blegdamsvej 3, DK-2200, Copenhagen, Denmark
   T. Moos & K. Møllgård

Authors

1. T. Moos
   You can also search for this author inPubMed Google Scholar
2. K. Møllgård
   You can also search for this author inPubMed Google Scholar

Rights and permissions

About this article

Cite this article

Moos, T., Møllgård, K. A sensitive post-DAB enhancement technique for demonstration of iron in the central nervous system.Histochemistry 99, 471–475 (1993). https://doi.org/10.1007/BF00274100

Download citation

• Accepted: 24 March 1993
• Issue Date: June 1993
• DOI: https://doi.org/10.1007/BF00274100

Keywords