DETERMINATION OF DEHYDROGENASE ACTIVITY IN TISSUE SECTIONS BY TRIDENSITOMETRY AND ITS APPLICATION (<Symposium>Multiparametric Photometry and Histo-and Cytochemistry)

概要

論文の詳細を見る
To compensate uneven section thickness in the cytophotometric determination of dehydrogenase activity demonstrated with nitroblue tetrazolium (NBT), the protein staining with naphthol yellow S (NYS) was superimposed and the amount of formazan was estimated as the ratio of tissue protein content instead of the direct correction of section thickness. The absorbance of superimposed section was measured at 700 nm, 570 nm and 450 nm for the background amount of formazan, formazan and tissue protein, respectively. Then, the enzyme activity was expressed as a relative activity (RA) as follows: RA=A570/(A450-A700) where, A570 and (A450-A570) represent the amounts of formazan and tissue protein, respectively. The quantitative reliability was tested with the polyacrylamide gel film containing mouse liver soluble fraction. For rapid and semi-automatic determination of RA, a special microphotometer, tridensity-automicrophotometer (TRIDENT), was devised. The method with TRIDENT was applied to show the quantitative distribution pattern of succinate dehydrogenase activity in various tissues. A quantitative topography has been revealed in various structural units of the liver lobule, gastric gland and muscle fiber for succinate dehydrogenase activity.
日本組織細胞化学会の論文