骨骼肌活组织检查病理诊断技术的改进及应用  

Improvement and application of diagnostic techniques of the skeletal muscle biopsy

作　　者：张燕[1] 韩志慧[1] 钟延丰[1] 王盛兰[1] 李玲玲[1] 郑丹枫[1] 

机构地区：[1]北京大学基础医学院病理学系,北京100191

出　　处：《北京大学学报（医学版）》2009年第4期459-462,共4页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(30670794)资助~~

摘　　要：目的:对300例骨骼肌活组织检查(活检)标本进行组织学切片和染色,总结分析技术方法的操作过程和改进,重点探讨各种肌酶及非酶组织化学染色技术在神经肌肉病诊断中的应用选择和价值。方法:将标本经异戊烷-液氮速冻后进行冰冻切片,应用HE及Gomori三色(GMR)、糖原(PAS)、脂肪(油红O)等特殊染色,肌球蛋白三磷酸腺苷酶(ATP酶)、还原型辅酶Ⅰ四唑氮还原酶(NADH-TR)等酶组织化学染色,并对各种染色技术进行调整和改进。结果:标本经异戊烷-液氮速冻及恒冷切片,可以有效避免产生冰晶。GMR染色主要显示线粒体及肌纤维变性坏死的病变,油红O染色显示肌纤维内有无脂质增多,PAS染色主要显示糖原和糖蛋白。对于肌肉组织,应用冰冻切片优于石蜡切片。NADH-TR及ATP酶染色可以区分两型肌纤维,并可观察肌纤维内部结构及线粒体酶的变化等。结论:通过骨骼肌酶与非酶组织化学染色技术,可以清晰地显示出两型肌纤维的分布情况及各种特征性的病理改变,几种方法可互为补充。Objective ： To summarize the improvement of various muscle staining techniques and discuss their application in diagnosis of neuromuscular diseases. Methods： Three hundred cases of skeletal muscle biopsy samples were examined by histopathological methods. The flash-freezing techniques were used for the preparation of frozen section, which were stained with HE, Gomori trichromie （GMR） , glycogen （PAS）, and fat acid （oil red O）; the enzyme histochemical staining with myosin adenosine triphos- phatase （ATPase）, and NADH-TR. Those stained methods had been improved. The immunohistoehemical staining with dystrophin; and transmission electronic microscopy were used. Results： Deepfreeze with heteropentane- liquid nitrogen and flash-freezing techniques could avoid artifacts of ice crystal vacuolation. GMR staining mainly showed the degenerative and necrotic lesions of mitochondria and muscle fibers. Oil red O staining showed the increase of lipid in muscle fibers. PAS staining mainly showed glycogen and gly- coprotein. Application of frozen sections in muscular tissue was better than that of paraffin sections. NADH-TR staining and ATPase staining could distinguish the two types of muscle fibers, and show the changes of inner structure of muscle fibers and mitochondria enzymes. Conclusion： The distribution and characteristic pathological changes of the two types of muscle fibers can be showed clearly by enzyme and non-enzyme histochemical staining techniques of the skeletal muscle. These methods can compliment with each other. Only after understanding the technical principles can we master and apply these methods.

关 键 词：肌 骨骼 活组织检查 冷冻超薄切片术 免疫组织化学 染色与标记 

分 类 号：R361.2[医药卫生—病理学]