显微曝光控制器对大鼠肾碱性磷酸酶组织化学进行定量研究的可行性探讨

A HISTOCHEMICAL STUDY OF ALKALINE PHOSPHATASE(AKP) DYNAMICS OF RAT RENAL TUBULE

采用大鼠肾皮质新鲜冰冻连续切片，厚６μｍ，对相邻切片进行不同时间、ｐＨ值和底物浓度的Ｇｏｍｏｒｉ钙—钴法显示碱性磷酸酶（ＡＫＰ）。反应后的标本用图像分析仪测试上述不同组织化学反应条件下肾小管ＡＫＰ反应产物的灰度值，进行定量分析。结果：①随着反应时间延长，灰度相应增加；②在ｐＨ７和ｐＨ１１条件下无反应产物，ｐＨ９和ｐＨ９．４时反应产物的灰度值最大；③随着底物浓度的增高，灰度值相应增加。根据上述实验结果绘制出不同时间、ｐＨ值、底物浓度的反应曲线，可得最佳反应时间、最适ｐＨ值和米氏常数。

Fresh renal cortex was sectioned by freezing microtome to serial sections at 6 ≥m.Adjacent sections were stained by means of Gomori Ca Co method at different time,different pH and different substrate concentrations to show alkaline phosphatase .The ash value of renal tubule AKP reactive products of all above different conditional reactions was measured by an automatic image analyzer and quantitative analysis was used.The results were as follows:①Along with the time prolonging,the ash value correspondingly increased.②There were no reactive products undre pH7 and pH11 conditions.The ash value of pH9 or pH9.4 was the largest.③Along with the substrate concentration becoming higher,the ash value correspondingly increased.According to the above results,the reactive curves at different time,different pH and different substrate concentrations were drawn.The most proper reactive time,the most suitable pH and Michaelis constant of AKP histochemistry of renal cortex fresh freezing section were obtaind.