Zur Methodik der elektronenmikroskopischen Darstellung der Succinat-Tetrazoliumsalzreduktase mit TC-NBT

Egger, G.

doi:10.1007/bf00306262

Cited by 5 publications

(1 citation statement)

References 6 publications

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“…BT (TC-NBT) (Seligman et aI., 1967;Boadle & Bloom, 1969;Egger, 1973) are osmiophilic, whereas the tetrazolium salts are non-osmiophilic. Therefore, these tetrazolium salts are suitable for electron microscopical studies (Seidler, 1991;Lewis & Knight, 1992).…”

Section: Reduction Methodsmentioning

confidence: 99%

In situ heterogeneity of peroxisomal oxidase activities: An update

Munchof

1996

Histochem J

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Oxidases are a widespread group of enzymes. They are present in numerous organisms and organs and in various tissues, cells, and subcellular compartments, such as mitochondria. An important source of oxidases, which is investigated and discussed in this study, are the (micro)peroxisomes. Oxidases share the ability to reduce molecular oxygen during oxidation of their substrate, yielding an oxidized product and hydrogen peroxide. Besides the hydrogen peroxide-catabolizing enzyme catalase, peroxisomes contain one or more hydrogen peroxide-generating oxidases, which participate in different metabolic pathways. During the last four decades, various methods have been developed and elaborated for the histochemical localization of the activities of these oxidases. These methods are based either on the reduction of soluble electron acceptors by oxidase activity or on the capture of hydrogen peroxide. Both methods yield a coloured and/or electron dense precipitate. The most reliable technique in peroxisomal oxidase histochemistry is the cerium salt capture method. This method is based on the direct capture of hydrogen peroxide by cerium ions to form a fine crystalline, insoluble, electron dense reaction product, cerium perhydroxide, which can be visualized for light microscopy with diaminobenzidine. With the use of this technique, it became clear that oxidase activities not only vary between different organisms, organs, and tissues, but that heterogeneity also exists between different cells and within cells, i.e. between individual peroxisomes. A literature review, and recent studies performed in our laboratory, show that peroxisomes are highly differentiated organelles with respect to the presence of active enzymes. This study gives an overview of the in situ distribution and heterogeneity of peroxisomal enzyme activities as detected by histochemical assays of the activities of catalase, and the peroxisomal oxidases D-amino acid oxidase, L-alpha-hydroxy acid oxidase, polyamine oxidase and uric acid oxidase.

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Section: Reduction Methodsmentioning

confidence: 99%

In situ heterogeneity of peroxisomal oxidase activities: An update

Munchof

1996

Histochem J

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Studies on the optimalisation and standardisation of the light microscopical succinate dehydrogenase histochemistry

Kügler

Wrobel

1978

Histochemistry

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The present investigation was undertaken in order to establish an optimal tissue pretreatment and an optimal incubation medium for the histochemical demonstration of succinate dehydrogenase (E.C. 1.3.99.1). The investigations were performed on steroid producing (testicle, adrenal gland) and steroid dependent (Fallopian tube) tissues. We studied the influences fo formalin fixation, acetone, magnesium ions, cyanides, electron carries (phenazine methosulfate, menadione coenzyme Q10), osmolarity, substrate concentration and inhibitors (oxalacetate, oxalate, malonate, 4-chloromercuribenzoic acid). The following procedure yields blameless morphological integrity and enzyme localization as well as optimal SDH-activity: Freezing of tissue cubes (diameter less than 5 mm) in propane cooled with liquid nitrogen or in melting freon. Incubation of 5 micrometer cryostat sections in narrow jars in the following medium (38.5 ml):--10 ml of 0.2 M sodium phosphate buffer pH 7.6 (52 mM).--18 mg tetranitro-BT in 0.5 ml dimethylformamide and aqua bidest. ad 10 ml (0.5 mM).--2.6 mg KCN in 16 ml aqua bidest. (1 mM).--540 mg succinate (disodium salt, hexahydrate) in 2 ml aqua bidest. (52 mM).--3 mg PMS (phenazine methosulfate) in 0.5 ml aqua bidest. (0.25 mM). The incubation medium has an osmolarity of 440 mosm. The incubation is carried out for 10 min at 37 degree C in darkness. To avoid non specific formazan deposits in lipid containing tissues a preincubation of the cryostat sections in 100% acetone at--22 degree C or--40 degree C for 7--10 min and an incubation time of 20--30 min is recommended. Control incubations adduced proof at the specificity of the SDH demonstration. Parallel incubation without PMS in order to determine indirectly the content of endogenous CoQ10 is further recommended.

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Electron Microscope Histochemistry

McClure

1981

Plant Parasitic Nematodes

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Zur Methodik der elektronenmikroskopischen Darstellung der Succinat-Tetrazoliumsalzreduktase mit TC-NBT

Cited by 5 publications

References 6 publications

In situ heterogeneity of peroxisomal oxidase activities: An update

In situ heterogeneity of peroxisomal oxidase activities: An update

Studies on the optimalisation and standardisation of the light microscopical succinate dehydrogenase histochemistry

Electron Microscope Histochemistry

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