摘要
目的:优化大鼠肺动脉平滑肌细胞(PASMCs)骨架蛋白F-actin图像的激光共聚焦显微技术。方法:采用不同浓度梯度的荧光染料及不同孵育条件,应用激光共聚焦显微扫描技术对PASMCs细胞骨架F-actin进行形态学观察。结果:终浓度10 mg/mL FITC-鬼笔环肽及10 mg/mL PI,分别染色30 min,并经RNA酶37℃处理为最适实验条件,可见F-actin呈明亮细丝状绿色荧光,PI标记的胞核呈红色荧光,胞浆无明显PI染色。结论:通过对染色条件的最终优化,表现出激光共聚焦技术在平面及三维同时观察细胞骨架F-actin的优越性,为进一步实验提供良好的技术平台。
Objective：To optimize the techniques of laser confocal scanning microscope（LCSM） on the image of F-actin in PASMCs.Method： Using different concentrations of fluorescent dyes and different incubation conditions,the image of F-actin in PASMCs was observed with CLSM.Results： Final concentration of 10 μg /mL FITC-Phalloidin,10 μg /mL PI,stained for 30 minutes,and worked by the Rnase at 37 ℃ were the optimal experimental conditions.The image showed that F-actin was filamentous green fluorescent light,PI labeled nuclei red fluorescence,and no significant PI staining cytoplasm.Conclusion： Through the final optimization of dyeing conditions,the experiment showed the superiority of the techniques of LCSM to observe F-actin in plane and three-dimensional cytoskeleton,which provided a good technical platform for further experiments.
出处
《电子显微学报》
CAS
CSCD
北大核心
2011年第1期61-64,共4页
Journal of Chinese Electron Microscopy Society
基金
浙江省中医药管理局资助项目(No.2010ZA084)
