CYTOPHOTOMETRY FOR SUCCINATE DEHYDROGENASE ACTIVITY IN RELATION TO TISSUE PROTEIN CONTENT WITH TETRANITRO BLUE TETRAZOLIUM (TNBT)

概要

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A relative cytophotometry was effectively applied for quantitative determination of succinate dehydrogenase (SDH) activity in rat liver and skeletal muscle section. The sections were stained first for SDH activity and then superimposed with protein staining by naphthol yellow S (NYS). The absorbance at 570 um and 450 um were measured cytophotometrically for the amount of TNBT-formazan and tissue protein content. The final result was expressed as a relative activity (RA) based on the ratio of the amount of TNBT-formazan to tissue protein content for the compensation of uneven section thickness. The linear relationship was obtained between the time of incubation and RA value of SDH in rat muscle section, as well as in rat liver section. In application, the present method has revealed the different frequency distribution profiles of RA of SDH in type I and II muscle fibers which were identified on the basis of the myofibrillar actomyosin ATPase activity. The RA value was less scattered in type I fiber than in type II fiber and the type II fiber could be subclassified into two groaps according to the RA-frequency profile for SDH in muscle fibers. This observation was almost in agreement with that in other studies on SDH activity-frequency profile for type II fiber. Thus, the present method was applicable to the cytophotometric determination of SDH activity in tissue sections.
日本組織細胞化学会の論文