ATP酶组织化学染色孵育方法比较

A comparison of incubation methods in ATPase histochemical staining

目的比较经典的ATP酶组织化学染色法中几种孵育方法优缺点,以用于临床诊断中肌纤维分型研究和肌肉疾病病理诊断。方法新鲜的肌肉组织用异戊烷-液氮冷冻,冰冻切片厚度7μm,组织化学染色采用ATP酶滴染法和浸染法两种,采用的两种孵育条件分别为37℃恒温箱(120~180)min和4℃冰箱过夜。结果 4℃过夜滴染的染色效果比37℃滴染的效果好;37℃浸染比4℃过夜浸染效果好。浸染过夜虽然容易有沉淀,但是使用前过滤ATP酶孵育液,染色效果仍然好。结论各种方法和实验条件的改变导致染色结果的不同,临床工作者可根据实际情况,在减少人力和试剂成本前提下,选择使用不同的ATP酶组织化学染色技术,使其得到更广泛的应用。

Objective To compare the incubation methods commonly used in ATPase histochemical staining for muscle fiber classification and the pathological diagnosis of muscle diseases.Methods Fresh muscle tissues were snap-frozen using isopento liquid nitrogen followed by sectioning w ith a thickness of 7 jun fo r each slice.Samples were subject to ATPase histochemical staining using two incubation methods of either dropping or dipping at 37℃fo r 2-3 hours or 4℃overnight.Results The dropping method gave a better dyeing effect at 4℃overnight,w hile dipping method worked better at 37℃.Conclusion The changes of methodology showed clear differences in the tissue staining effects,which allows clinicians more fle xib ility to select appropriate method,thus broadening the application of ATPase histochemistry staining technique w ith reduced labor and reagent costs.