不同冻存法对肌肉冰冻切片后HE染色和肌球蛋白ATP酶染色效果的影响

Effects of Different Cryopreserving Methods on the Results of HE Staining and Myosin ATP Staning of Frozen Sections of Longissimus Dorsi

使用分步法和一步法冻存巴马香猪背最长肌,冰冻切片后比较苏木精-伊红染色法(HE染色)和肌球蛋白三磷酸腺苷酶染色法(ATP酶染色)效果。结果表明:分步法冻存的肌肉样品冰晶少,细胞形态完整,HE染色效果好,核质明显;ATP酶染色效果好,可较好地区分Ⅰ型Ⅱ型肌纤维。一步法冻存的肌肉样品冰晶多,细胞形态不完整,HE染色效果差,核质模糊;ATP酶染色效果差,基本染不上色,无法对肌纤维类型进行区分。分步冻存法的建立为通量式制备猪肉的恒冻切片奠定了基础,也为进一步开展相关性状的生理学、遗传学及种猪选育提供了很好的表型测定方法。

In this study,the results of hematoxylin eosin staining（ HE staining） and myosin ATPase staining were compared by step-by-step method and one-step method cryopreserving longissimus dorsi muscle of Bama Xiang pig.The results showed that the samples cryopreserved by step-by-step method contained less ice crystals,the cells were complete,the effect of HE staining was very good in which nucleus and cytoplasm could be observed clearly and the effect of ATP enzyme staining was good enough in which type I and type II muscle fibers could be distinguished easily; the samples cryopreserved by one-step method contained a large amount of ice crystals cells were broken,the effect of HE staining was very poor in which nucleus and cytoplasm could not be observed,and type I and type II muscle fibers could not be distinguished since myosin ATPase could hardly be stained on the frozen sections.The establishment of step-by-step cryopreserving method laid the foundation for preparing large number of frozen sections of pork and provided a good way for penotype indentification for further analysis of physiology,genetics and breeding.