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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 frozen sections of rat renal cortex were used to measure the alkaline phosphatase (AKP) by Gomori calcium - cobalt method. After the reaction, the image analyzer was used to test the gray value of the renal tubular AKP reaction product under the above-mentioned different tissue chemical reaction conditions for quantitative analysis. Results: ① With the reaction time prolonged, the gray level increased accordingly. ② The reaction products had the highest gray value at pH7 and pH11, pH9 and pH9.4. With the increase of substrate concentration, The value increases accordingly. According to the above experimental results, the reaction curves of different time, pH value and substrate concentration were drawn, and the optimum reaction time, optimum pH value and Michaelis constant were obtained.