Enhanced light microscopic visualization of oxidase activity with the cerium capture method

Abstract

Visualization methods for the light microscopic detection of the activity of oxidases after being localized with cerium ions as reported by Angermüller and Fahimi (1988a, b) are not suitable for the demonstration of H2O2-genrating oxidases at sites with low activity. Therefore, the cerium-diaminobenzidine (DAB) visualization procedure of these authors was modified. Nickel or cobalt ions were added to the DAB solution together with small amounts of H2O2. Visualization was performed in a one-step-method. This modified visualization technique enables light microscopic detection of amino acid oxidase activity in kidney and liver cells where it was found with the original method but the amounts of final reaction product were considerably higher. Moreover, the DAB-nickel-H2O2 and DAB-cobalt-H2O2 procedures were more sensitive than the cerium-lead method of Angermüller and Fahimi (1988a, b). The method appeared to be specific, because final reaction product was not found after control incubation. Especially the DAB-nickel-H2O2 procedure can also be used for immunohistochemistry when glucose oxidase serves as the enzyme label.