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Nichtenzymatische TNBT-Färbung von Gewebsstrukturen

Licht- und elektronenmikroskopische Untersuchungen

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Summary

Incubating native or fixed cryostat sections in a TNBT-containing medium and reducing TNBT to formazan, produces a non-enzymatic staining of tissue structures. This phenomenon is investigated light and electron microscopically; the emphasis is put on the intracellular localization of TNBT-formazan. In the cell electron-dense TNBT-formazan is found in the immediate vicinity of the external membranes of mitochondria, membranes of the endoplasmic reticulum, and in regions, which are free from organelles. The reaction becomes weaker if it is preceded by an extraction of lipids. It is therefore concluded that TNBT is absorbed non-specifically onto the lipoproteins of the cell and that the localization of TNBT-formazan is influenced by diffusion phenomenons. It seems possible that TNBT is reduced by free diffusible reducing tissue components. If TNBT is used as an indicator for reduction in light or submicroscopical investigations of oxidative enzymes a non-specific TNBT-formazan localization in tissue structures might be encountered. However, more work along these lines is needed to finally confirm this theory.

Zusammenfassung

Eine Inkubation nativer oder fixierter Kryostatschnitte in einem TNBT-haltigen Inkubationsmedium und eine anschließende Reduktion des TNBT zum Formazan führt zu einer nichtenzymatischen Anfärbung der Gewebsstrukturen. Diese Beobachtung wird licht- und elektronenmikroskopisch im Hinblick auf eine intrazelluläre Lokalisation des TNBT-Formazans näher analysiert. Elektronendichtes TNBT-Formazan konnte in der Zelle in unmittelbarer Nachbarschaft von Mitochondrienaußenmembranen, Membranen des endoplasmatischen Retikulums und in organellenfreien Bezirken nachgewiesen werden. Eine vorherige Lipidextraktion bewirkt eine Abschwächung der Reaktion. Es wird daraus der Schluß gezogen, daß TNBT unspezifisch an Lipoproteinstrukturen der Zelle adsorbiert wird und daß die beobachtete Lage des TNBT-Formazans durch Diffusionsphänomene beeinflußt wird. Möglicherweise kann das TNBT durch frei diffusible reduzierende Gewebskomponenten reduziert werden. Bei Verwendung von TNBT als Reduktionsindikator zum licht- und submikroskopischen Nachweis oxydativer Enzyme ist mit einer unspezifischen TNBT-Formazan-Lokalisation in Gewebsstrukturen zu rechnen. Die endgültige Bestätigung dieser Auffassung erfordert weitere Modellversuche.

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Authors and Affiliations

  1. Pathologisches Institut der Karl-Marx-Universität Leipzig, Deutschland

    F. Wohlrab & U. Fuchs

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  1. F. Wohlrab
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  2. U. Fuchs
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Wohlrab, F., Fuchs, U. Nichtenzymatische TNBT-Färbung von Gewebsstrukturen. Histochemie 9, 256–268 (1967). https://doi.org/10.1007/BF00294393

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  • DOI: https://doi.org/10.1007/BF00294393

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