Murray G I, Burke M D, Ewen S W
Department of Pathology, University of Aberdeen, UK.
Histochem J. 1988 Sep;20(9):491-8. doi: 10.1007/BF01002647.
A method has been developed for the histochemical demonstration of a variety of dehydrogenases in freeze-dried or fixed resin-embedded tissue. Seven dehydrogenases were studied. Lactate dehydrogenase, NADH dehydrogenase and NADPH tetrazolium reductase were all demonstrable in sections of paraformaldehyde-fixed resin-embedded tissue. Freeze-dried specimens were embedded, without fixation, in glycol methacrylate resin or LR Gold resin at either 4 degrees C or -20 degrees C. All the dehydrogenases except succinate dehydrogenase retained their activity in freeze-dried, resin-embedded tissue. Enzyme activity was maximally preserved by embedding the freeze-dried tissue specimens in glycol methacrylate resin at -20 degrees C. The dehydrogenases were accurately localized without any diffusion when the tissue sections were incubated in aqueous media. Addition of a colloid stabilizer to the incubating medium was not required. Freeze-drying combined with low-temperature resin embedding permits accurate enzyme localization without diffusion, maintenance of enzyme activity and excellent tissue morphology.
已开发出一种用于在冻干或固定树脂包埋组织中进行多种脱氢酶组织化学显示的方法。研究了七种脱氢酶。乳酸脱氢酶、NADH脱氢酶和NADPH四氮唑还原酶在多聚甲醛固定树脂包埋组织切片中均能显示。冻干标本在4℃或-20℃下不经过固定，直接包埋于甲基丙烯酸乙二醇酯树脂或LR Gold树脂中。除琥珀酸脱氢酶外，所有脱氢酶在冻干树脂包埋组织中均保持其活性。将冻干组织标本在-20℃下包埋于甲基丙烯酸乙二醇酯树脂中，酶活性得到最大程度的保留。当组织切片在水性介质中孵育时，脱氢酶能准确定位且无任何扩散。孵育介质中无需添加胶体稳定剂。冻干结合低温树脂包埋可实现酶的准确定位而无扩散、保持酶活性并获得优良的组织形态。
