The cerium perhydroxide-diaminobenzidine (Ce-H2O2-DAB) procedure |
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| Authors: | K -J Halbhuber R Gossrau U Möller C E Hulstaert N Zimmermann H Feuerstein |
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| Institution: | (1) Institute of Anatomy, Department of Histochemistry, Friedrich Schiller University Jena, Teichgraben 7, DDR-6900 Jena, German Democratic Republic;(2) Department of Anatomy, Free University of Berlin, Königin-Luise-Strasse 15, D-1000 Berlin 33, Germany;(3) Centre for Medical Electron Microscopy, University of Groningen, Oostersingel 69/2, Groningen, The Netherlands |
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| Abstract: | Summary New light microscopic visualization methods were developed for the histochemical detection of non-specific alkaline and acid phosphatase, Mg-, Ca-and Na, K-dependent adenosine triphosphatase, myosin adenosine triphosphatase, glucose-6-phosphatase, 5 -nucleotidase and thiamine pyrophosphatase with cerium ions as trapping agents in cryostat and plastic sections. The techniques are based on the conversion of cerium phosphate into cerium perhydroxide by H2O2 which decomposes at 55°–60° C into cerium hydroxide and oxygen radicals. These radicals are able to oxidize diaminobenzidine (DAB) to DAB brown. Addition of nickel ions to the DAB-H2O2 mixture generates bluish-black stained nickel-DAB complexes. Compared with the classical metal precipitation, azo, azoindoxyl and tetrazolium procedures the H2O2-DAB and especially the H2O2-DAB-nickel methods provided identical or superior results in catalytic phosphatase histochemistry and immunohistochemistry when using non-specific alkaline phosphatase as the enzyme label. |
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