改良的肥大细胞甲苯胺蓝染色法  

The Modified Toluidine Blue Staining Method for Mast Cell

作　　者：梁玉婷[1] 彭霞[1] 林堃[1] 尹悦[1] 李莉[1] 

机构地区：[1]上海交通大学附属第一人民医院检验科,上海200080

出　　处：《现代生物医学进展》2017年第24期4601-4605,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81601395);上海市科委引导类项目(14411963200);(16YF1409200)

摘　　要：目的:本研究旨在运用一种改良的甲苯胺蓝染色方法鉴定体外分离培养的肥大细胞形态,为肥大细胞研究提供简便的检测方法。方法:体外诱导分化培养小鼠骨髓来源的肥大细胞,4周后收集细胞、固定、染色。比较不同固定温度及时间对甲苯胺蓝染色效果的影响,确定最佳条件。结果:SCF和IL-3体外培养诱导出小鼠骨髓来源肥大细胞。肥大细胞用改良的甲苯胺蓝染色后细胞着色效果较好,细胞多呈圆形或椭圆形且胞膜较完整,胞浆中充满大量的紫红色颗粒。结论:本研究成功运用一种适用于体外培养小鼠骨髓源肥大细胞的甲苯胺蓝染色法,并发现肥大细胞在37℃充分固定后进行染色,可以降低肥大细胞发生脱颗粒。此操作方法稳定性好、简便,适用于体外培养的肥大细胞形态学观察。Objective：This study aimed to identify the morphology of mast cells by using a modified toluidine blue staining scheme,so as to provide a powerful reference for the experimental basis research of mast cells.Methods：Bone marrow-derived mast cells were induced in vitro.After 4 weeks,the cells were collected,fixed,and stained.Mast cells were fixed at different temperature during different time.The optimum condition was determined by comparing the effects of toluidine blue staining.Results：Bone marrow cells were induced to differentiate into mast cells by SCF and IL-3 in vitro.When mast cells were stained with modified toluidine blue staining,the staining effect was better.Mast cells were round or oval and the cell membrane was complete and the cytoplasm was filled with a large number of purple particles.Conclusion：In this study,we successfully applied a modified toluidine blue staining method to mast cells cultured in vitro.The results showed that the condition at 37 ℃ full fixation with staining could reduce the degeneration of mast cells.This method was easy to operate with good stability.It was suitable for the morphological observation of mast cells cultured in vitro.

关 键 词：肥大细胞 甲苯胺蓝 染色 

分 类 号：R-33[医药卫生] R446.61