小鼠肝脏弹性蛋白免疫荧光染色方法优化

摘要

Objective To explore the improved method of elastin (ELN)immunofluorescent staining in examination of mice liver,and to provide the technical basis for studying the deposition and degradation of ELN. Methods The frozen liver sections of male C57BL/ 6 mice were used for routine or improved immunofluorescent staining. In comparison to the routine method,additional incubation with hyaluronidase (Haase) or collagenase (Colase)or both were manipulated before the performation of electron microscopy for immune - labelled ELN. Results No signifi-cantly positive ELN area had been detected in the liver with both conventional staining group and Colase incubated group. In Haase incubated group,some discontinuous ELN - positive staining could be seen in liver sections. The positive staining area of ELN was significantly increased af-ter Haase and Colase combined incubation,mainly distributed in the areas of hepatic artery,portal vein and extracellular matrix. Conclusion Combined incubation of Haase and Colase before the formation of ELN antibody binding protein can significantly raise the effect of staining in ELN immunofluorescent staining of mice liver.%目的 探讨肝脏弹性蛋白(ELN)免疫荧光染色方法 的优化,改善肝脏ELN染色效果.方法小鼠肝脏冰冻切片,分别进行常规和优化的免疫荧光染色.优化方法分别是ELN抗体结合蛋白前孵育透明质酸酶(HAase)或/和胶原酶(COLase).观察小鼠肝脏ELN免疫荧光染色效果,并统计ELN阳性面积与肝脏面积比值.结果 小鼠肝脏冰冻切片经常规和孵育COLase免疫荧光染色均未见明显ELN阳性面积.孵育HAase可在肝脏血管部位见少量不连续的ELN阳性着色.联合孵育HAase和COLase后ELN阳性面积明显增加,主要见于肝动脉、门静脉和周围的细胞间质内.结论 HAase和COLase联合孵育可显著改善小鼠肝脏ELN免疫荧光染色效果.