Abstract
Peroxisomes are ubiquitous dynamic and multifunctional organelles that contribute to numerous anabolic and catabolic pathways, being essential for human health and development. Their best known functions include the oxidation of fatty acids and metabolism of hydrogen peroxide with catalase as a marker enzyme. Indeed, historically, it was the cytochemical staining of catalase in many different cells and tissues that revealed the ubiquitous presence of peroxisomes in almost all animal and plant cells. In this chapter, the method for cytochemical staining of catalase with the alkaline 3, 3′-diaminobenzidine (DAB) is described. Since aldehyde fixation is a prerequisite for staining of catalase with DAB, a method for perfusion fixation of rat liver with glutaraldehyde is presented prior to the cytochemical staining method and the subsequent tissue processing for light and electron microscopy.
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References
Rhodin J (1954) Correlation of ultrastructural organization and function in normal and experimentally changed proximal convoluted tubule cells of the mouse kidney. Doctorate Thesis. Karolinska Institute, Stockholm
Rouiller C, Bernhard W (1956) “Microbodies” and the problem of mitochondrial regeneration in liver cells. J Biophys Biochem Cytol 2(Suppl):355–359
De Duve C, Baudhuin P (1966) Peroxisomes (microbodies and related particles). Physiol Rev 46:323–357
Fahimi HD (1968) Cytochemical localization of peroxidase activity in rat hepatic microbodies (peroxisomes). J Histochem Cytochem 16:547–550
Fahimi HD (1969) Cytochemical localization of catalase in rat hepatic microbodies (peroxisomes). J Cell Biol 43:275–288
Hruban Z, Vigil EL, Slesers A, Hopkins E (1972) Microbodies: constituent organelles in animal cells. Lab Invest 27:184–191
Angermüller S, Fahimi HD (1981) Selective cytochemical localization of peroxidase, cytochrome oxidase and catalase in rat liver with 3,3′-diaminobenzidine. Histochemistry 71:33–44
Bonekamp NA, Islinger M, Lazaro MG, Schrader M (2013) Cytochemical detection of peroxisomes and mitochondria. Methods Mol Biol 931:467–482
Völkl A, Fahimi HD (1985) Isolation and characterization of peroxisomes from the liver of normal untreated rats. Eur J Biochem 149:257–265
Teorell I, Stenhagen E (1938) Ein Universalpuffer für den pH-Bereich 2,0-12,0. Biochem Z 299:416–419
Herzog V, Fahimi HD (1974) The effect of glutaraldehyde on catalase. Biochemical and cytochemical studies with beef liver catalase and rat liver peroxisomes. J Cell Biol 60:303–311
Yamamoto K, Völkl A, Hashimoto T, Fahimi HD (1988) Catalase in guinea pig hepatocytes is localized in cytoplasm, nuclear matrix and peroxisomes. Eur J Cell Biol 46:129–135
Fahimi HD (1974) Effect of buffer storage on fine structure and catalase cytochemistry of peroxisomes. J Cell Biol 63:675–783
Fahimi HD (1973) Diffusion artifacts in cytochemistry of catalase. J Histochem Cytochem 21:999–1009
Legg PG, Wood RL (1970) New observations on microbodies. A cytochemical study on CPIB treated rat liver. J Cell Biol 45:118–129
Yokota S, Oda T, Fahimi HD (2001) The role of 15-lipoxygenase in disruption of the peroxisomal membrane and in programmed degradation of peroxisomes in normal rat liver. J Histochem Cytochem 49:613–622
Fahimi H D (1975) Fine-Structural cytochemical localization of peroxidatic activity of catalase. Tech Biochem Biophys Morphol (Glick D, Rosenbaum R M ed.) 2:197–245
Acknowledgments
It is a pleasure to thank all my former associates and colleagues for many years of fruitful and stimulating discussions. For technical support I would like to thank Gabi Krämer and Inge Frommer and for logistics Annemarie Achten, companions of many years. My work in USA was supported by grants from NIH and in Germany from DFG. In addition, we were supported by research funds of Boehringer Mannheim (now Roche Scientific).
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Fahimi, H.D. (2017). Cytochemical Detection of Peroxisomes in Light and Electron Microscopy with 3,3′-diaminobenzidine. In: Schrader, M. (eds) Peroxisomes. Methods in Molecular Biology, vol 1595. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6937-1_10
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DOI: https://doi.org/10.1007/978-1-4939-6937-1_10
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