Light microscopic immunocytochemical demonstration of peroxisomal enzymes in epon sections |
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| Authors: | J A Litwin S Yokota T Hashimoto H D Fahimi |
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| Institution: | (1) Department of Anatomy, II. Division, University of Heidelberg, Im Neuenheimer Feld 307, D-6900 Heidelberg, Federal Republic of Germany;(2) Department of Anatomy, Yamanashi Medical School, Yamanashi, Japan;(3) Department of Biochemistry, Shinshu University, Matsumoto, Nagano, Japan;(4) Present address: Department of Histology, Institute of Biomorphology, Copernicus Medical Academy, Krakow, Poland |
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| Abstract: | Summary A procedure is described for light microscopic immunocytochemical localization of catalase and three enzymes of peroxisomal lipid  -oxidation: acyl-CoA oxidase, enoyl-CoA hydratase and 3-ketoacyl-CoA thiolase in semithin sections of rat liver processed for routine electron microscopy. Satisfactory immunostaining required the removal of the epoxy resin with sodium ethoxide, controlled digestion of deplasticized sections with proteases and, in case of osmiumfixed tissue, bleaching with oxidants. Resin removal was essential for successful immunostaining, and protease treatment enhanced markedly the intensity of the reaction. This study shows that tissues processed for conventional ultrastructural studies can be used for postembedding immunocytochemical demonstration of various peroxisomal enzymes.Supported by the Alexander-von-Humboldt Foundation |
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