Cobalt acetylacetonate-diaminobenzidine reaction for microspectrophotometry of cytochrome oxidase.

Hiraoka, Toshisuke; Hirai, Kei-Ichi; Ueda, Tadashi

doi:10.1267/ahc.19.429

Cited by 7 publications

(2 citation statements)

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“…The localization of final reaction product that was induced by addition of cytochrome c was similar to the localization described in the literature (Hiraoka et al, 1986) but differed from the localization after DAB-Mn-Co incubation in the absence of cytochrome c. The amount of final reaction product of cytochrome c oxidase activity was higher and was found in more cells than after incubation with the DAB-Mn-Co medium. Endogenous cytochrome c may be held responsible for the spontaneous formation of final reaction product.…”

Section: Discussionsupporting

confidence: 76%

“…These results indicate that NADH:coenzyme Q reductase and aldehyde oxidase are possible candidates (Kehrer, 1993). The ultrastructural localization of final reaction product at inner and outer membranes of mitochondria and at the intermembrane space also support the conclusion that the proposed enzymes are involved (Hiraoka et al, 1986;Critchley et al, 1992).…”

Section: Discussionsupporting

confidence: 52%

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KERVER

1997

The Histochemical Journal

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In situ detection of spontaneous superoxide anion and singlet oxygen production by mitochondria in rat liver and small intestine Kerver, E.D.; Vogels, I.M.C.; Bosch, K.S.; Vreeling-Sindelárová, H.; van den Munckhof, R.J.M.; Frederiks, W.M. Published in:The histochemical journal DOI:10.1023/A:1026453926517 Link to publication Citation for published version (APA):Kerver, E. D., Vogels, I. M. C., Bosch, K. S., Vreeling-Sindelárová, H., van den Munckhof, R. J. M., & Frederiks, W. M. (1997). In situ detection of spontaneous superoxide anion and singlet oxygen production by mitochondria in rat liver and small intestine. The histochemical journal, 29, 229-237. DOI: 10.1023/A:1026453926517 General rightsIt is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. Download date: 11 May 2018In situ detection of spontaneous superoxide anion and singlet oxygen production by mitochondria in rat liver and small intestine SummaryIn the present study, the endogenous formation of reactive oxygen species was localized in rat liver and small intestine. The 3,39-diaminobenzidine (DAB)-Mn 2 technique in which cobalt ions were included in the incubation medium was applied to unfixed cryostat sections of intact tissues. Addition of manganese ions to the DAB-Co 2 -containing medium greatly increased the amounts of final reaction product formed compared with incubations with only DAB and cobalt ions. In liver, a blue final reaction product was deposited, particularly in hepatocytes surrounding portal tracts. In the small intestine, the DAB-cobalt complex was mainly found at the basal side of enterocytes. Goblet cells remained unstained. Electron microscopical images revealed that an electron-dense reaction product was exclusively present at both inner and outer membranes and at the intermembrane space in mitochondria of liver parenchymal cells and duodenal enterocytes. It was shown that the spontaneous formation of final reaction product was enzymatic and dependent on the presence of oxygen in the medium. Sulphide decreased the reaction, which may indicate that cytochrome c oxidase was partially involved. Benzoquinone and histidine, which are scavengers of superoxide anions and singlet oxygen respectively, reduced the amount of final reaction product considerably. Furthermore, the formation of final reaction pro...

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Section: Discussionsupporting

confidence: 76%

Section: Discussionsupporting

confidence: 52%

Untitled

KERVER

1997

The Histochemical Journal

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Reflectance enzyme histochemistry (REH): visualization of cerium-based and DAB primary reaction products of phosphatases and oxidases in cryostat sections by confocal laser scanning microscopy

Scheven

Jirikowski

et al. 1996

Histochem Cell Biol

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In the present study the reflectance mode of confocal laser scanning microscopy was adapted to detect and to assess semiquantitatively cerium-based primary reaction products of oxidases [Ce(IV) perhydroxide] and phosphatases [Ce(III) hydroxyphosphate converted into Ce(IV) perhydroxyphosphate] as well as of the 3,3'-diaminobenzidine (DAB)-based primary reaction product of cytochrome c oxidase in cryostat sections. Confocal laser scanning microscopy offers a unique way of making visible histochemical reaction products which are weakly absorbant but sufficiently reflective. It was easily possible to record simultaneously the reflectance signals at the wavelength of the exciting laser (preferentially 488 nm) and the autofluorescence signals ( > 580 nm in our set-up) of glutaraldehyde-fixed tissue. The results of an imbibition study of cerium-containing model precipitates indicate that the cerium, generally, should be oxidized prior to observation because the index of refraction of Ce(IV) compounds is considerably higher than that of the corresponding Ce(III) compounds. An attempt at comparative numerical assessment of reflection intensities from reflectant parts in morphologically similar sections is presented. The proposed technique may open new possibilities in enzyme- and immunohistochemistry.

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Detection of endogenous and immuno-bound peroxidase — The status Quo in histochemistry

Krieg

2010

Progress in Histochemistry and Cytochemistry

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Cobalt acetylacetonate-diaminobenzidine reaction for microspectrophotometry of cytochrome oxidase.

Cited by 7 publications

References 10 publications

Untitled

Untitled

Reflectance enzyme histochemistry (REH): visualization of cerium-based and DAB primary reaction products of phosphatases and oxidases in cryostat sections by confocal laser scanning microscopy

Detection of endogenous and immuno-bound peroxidase — The status Quo in histochemistry

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