AMeX- 처리 파라핀절편을 이용한 alkaline phosphatase의 검출에 관한 조직화학적 연구

Abstract

The purpose of this study was performed to evaluate the usefulness of the acetonefixed, methyl benzoate and xylene cleared (AMeX -prepared) paraffin sections for demonstration of alkaline phosphatase(ALPase) activity. Tissues were fixed in acetone at - 20℃overnight, then cleared in methyl benzoate and xylene, consecutively, and embedded in ordinary paraffin at 58~60℃. Paraffin sections were deparaffinized with xylene, rehydrated with a descending graded alcohol series and then washed in distilled water, and stained with azo dye coupling method, which have generally been used on freshfrozen or cold acetone-fixed frozen sections. As with cold-acetone-fixed frozen sec tions, ALPase activity was clearly localized in the all tissue sections prepared with the present method : stomach, small intestine, liver, spleen, pancreas, kidney, adrenal, uterus, salpinx, ovary, heart, trachea and lung. High concentrations of ALPase, especially, were found in the brush borders of the intestinal mucosa and the proximal tubules of the kidney. ALPase was always observed in the capillary endothelium of all tissue sections, and also in the chief cells of stomach, eosinophilic leukocytes in intestinal mucosa, Kupffer cells of liver, reticular cells of spleen, basal cells and some surface epithelial cells of tracheal epithelium and alveolar macrophages of lung. In addition, isozyme patterns of ALPase in the all tissue sections could be classsified to two species, intestinal - type and other nonspecific type, based on response to 2.5 mM L -levamisole and 65℃ - pretreatment. These findings indicate that sections prepared by the present method demonstrated much better histologic and cytologic preservation than possible in frozen sections. Therefore, azo dye coupling method using AMeX - prepared paraffin sections is considered to be a useful method for demonstration of ALPase activity in rat tissues.