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Perfusion-Perls and -Turnbull methods supplemented by DAB intensification for nonheme iron histochemistry: demonstration of the superior sensitivity of the methods in the liver, spleen, and stomach of the rat

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Abstract

Perfusion-Perls and -Turnbull methods supplemented by the intensification with 3,3′-diaminobenzidine (+ DAB) enabled stronger and more extensive staining of nonheme iron than the Perls + and Turnbull + DAB methods carried out on tissue sections fixed with 10% formalin in 0.9% saline or PBS. The section- and perfusion-Perls + DAB methods are not specific for the demonstration of nonheme ferric iron but also stain nonheme ferrous iron. However, owing to its high sensitivity, the perfusion-Perls + DAB method would provide useful information about nonheme iron deposition regardless of oxidation states in normal and pathological conditions. The perfusion-Turnbull + DAB method is specifically demonstrable of nonheme ferrous iron and the results from this method showed significant stores of nonheme ferrous iron in the hepatocytes, Kupffer cells, splenic macrophages, and gastric parietal cells of the rat. Since nonheme ferrous iron is considered to be critically involved in free radical generation, the perfusion-Turnbull + DAB method would visualize such populations of cells that are at risk from free radical damage.

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Acknowledgements

This work was supported by Grant-in-Aid (14657123) from the Ministry of Education, Science, Sports and Culture, Japan, and by a grant from Aomori Bank, Aomori, Japan, for the research project "Cerebrovascular Disorders".

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Authors and Affiliations

  1. Department of Anatomy, Hirosaki University School of Medicine, 036-8562, Hirosaki, Japan

    Reiko Meguro, Yoshiya Asano, Hiroyasu Iwatsuki & Kazuhiko Shoumura

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  1. Reiko Meguro
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  2. Yoshiya Asano
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  3. Hiroyasu Iwatsuki
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  4. Kazuhiko Shoumura
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Correspondence to Reiko Meguro.

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Meguro, R., Asano, Y., Iwatsuki, H. et al. Perfusion-Perls and -Turnbull methods supplemented by DAB intensification for nonheme iron histochemistry: demonstration of the superior sensitivity of the methods in the liver, spleen, and stomach of the rat. Histochem Cell Biol 120, 73–82 (2003). https://doi.org/10.1007/s00418-003-0539-y

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