【摘要】 介绍如何利用ATP酶组织化学染色法测定骨骼肌的快、慢肌纤维类型。成年SD大鼠若干,断头处死,取后肢腓肠肌内侧头,立即进行液氮速冻切片法切片(厚度10μm),室温干燥30～60 min后,分别在酸预孵和碱预孵条件下进行ATP酶组织化学染色。结果显示,碱预孵(pH 10.4)后,Ⅱ型肌纤维被染成深褐色,Ⅰ型肌纤维则着色较浅;与此相反,经酸预孵(pH 4.3)后,Ⅰ型肌纤维被染成深褐色,Ⅱ型肌纤维则着色较浅。因此,无论是通过酸预孵(pH 4.35)还是碱预孵(pH 10.4)进行ATP酶组织化学染色都可以实现对快、慢肌纤维类型的测定。此实验不但可以加强学生对肌纤维类型的认识,还让学生接触了当前生命科学研究中常用的组织化学检测技术,并且所需的试剂与耗材相对便宜,具有一定的推广和使用价值。更多还原

【Abstract】 Using ATPase staining to identify the skeletal muscle fiber types,slow-twitch type(typeⅠ) and fast-twitch type(typeⅡ).Several adult male Sprague-Dawley(S-D) rats were euthanized,and their medial gastrocnemii of left hind limbs were collected immediately and processed into frozen cross sections at a thickness of 10 μm in a freezing cryostat at-20℃.The two types of muscle fibers were identified after staining for myofibrillar ATPase.After alkaline pre-incubation(pH 10.4),the type Ⅱ muscle fibers were stained to dark brown,the typeⅠjust stained slightly;on the contrary,after acid pre-incubation(pH 4.3),the type Ⅰ muscle fibers were stained to dark brown,the typeⅡ just stained slightly.The results demonstrate,no matter using an acid pre-incubation(pH 4.35)or an alkaline pre-incubation(pH 10.4),both kinds of ATPase staining can identify the types of skeletal muscle fibers.This experiment not only makes students understand the types of skeletal muscle fibers better,but also let them contact the histochemical staining technique,which is widely used in life scientific studies currently.The cost of the reagents and consumptive materials in this experiment is not expensive,so it is worth using and spreading.更多还原