title: Nonheme-iron histochemistry for light and electron microscopy: a historical, theoretical and technical review.
abstract: We reviewed the methods of nonheme-iron histochemistry with special focus on the underlying chemical principles. The term nonheme-iron includes heterogeneous species of iron complexes where iron is more loosely bound to low-molecular weight organic bases and proteins than that of heme (iron-protoporphyrin complex). Nonheme-iron is liberated in dilute acid solutions and available for conventional histochemistry by the Perls and Turnbull and other methods using iron chelators, which depend on the production of insoluble iron compounds. Treatment with strong oxidative agents is required for the liberation of heme-iron, which therefore is not stained by conventional histochemistry. The Perls method most commonly used in laboratory investigations largely stains ferric iron, but stains some ferrous iron as well, while the Turnbull method is specific for the latter. Although the Turnbull method performed on sections fails in staining ferrous iron or stains only such parts of the tissue where iron is heavily accumulated, an in vivo perfusion-Turnbull method demonstrated the ubiquitous distribution of ferrous iron, particularly in lysosomes. The Perls or Turnbull reaction is enhanced by DAB/silver/gold methods for electron microscopy. The iron sulfide method and the staining of redox-active iron with H(2)O(2) and DAB are also applicable for electron microscopy. Although the above histochemical methods have advantages for visualizing iron by conventional light and electron microscopy, the quantitative estimation of iron is not easy. Recent methods depending on the quenching of fluorescent divalent metal indicators by Fe(2+) and dequenching by divalent metal chelators have enabled the quantitative estimation of chelatable Fe(2+) in isolated viable cells.
authors.affiliation: Department of Anatomy, Hirosaki University School of Medicine, Hirosaki, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Meguro
authors.forename: Yoshiya
authors.initials: Y
authors.lastname: Asano
authors.forename: Saori
authors.initials: S
authors.lastname: Odagiri
authors.forename: Chengtai
authors.initials: C
authors.lastname: Li
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Iwatsuki
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Shoumura
mesh_terms: D000818:Animals; D005290:Ferric Compounds / Q000032:analysis / Q000378:metabolism; D005296:Ferrous Compounds / Q000032:analysis / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods*; D049672:History, 19th Century; D049673:History, 20th Century; D049674:History, 21st Century; D006801:Humans; D007501:Iron / Q000032:analysis / Q000266:history / Q000378:metabolism; D008853:Microscopy / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D019159:Nonheme Iron Proteins / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005290:Ferric Compounds; D005296:Ferrous Compounds; D019159:Nonheme Iron Proteins; D007501:Iron