全内反射双通道观察双标记荧光染色的非洲绿猴肾细胞  

Observation of Double-Stained African Green Monkey Kidney COS-7 Cells Using Total Internal Reflection Double-Channel Fluorescence Microscopy

作　　者：刘晓晨[1] 关立照[2] 马页云[1] 张宏权[2] 

机构地区：[1]清华大学物理系,原子分子纳米科学教育部重点实验室,北京100084 [2]北京大学医学部人体解剖与组织胚胎学系分子细胞生物学与肿瘤生物学实验室,北京100191

出　　处：《光谱学与光谱分析》2010年第10期2676-2679,共4页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(10874099,90919012,30830048);教育部博士点基金项目(20090002110065)资助

摘　　要：在像增强型电荷耦合器件(ICCD)荧光显微成像装置上用双通道成像方法观察了非洲绿猴肾细胞(COS-7)中由EGFP转染的肌球蛋白Myosin 15a,以及Rhodamine标记的细胞微丝。为观察微丝尖端的肌球蛋白Myosin 15a,采用了高灵敏、低损伤的全内反射激发荧光显微成像技术,并在双通道中选用合适滤光片组合消除两种荧光染料间的光谱串扰。实验观测到非洲绿猴肾细胞内过表达的肌球蛋白Myosin 15a和伸长的微丝的分布情况,尤其是清晰观察到Myosin 15a在微丝上的分布。为全内反射双通道荧光成像技术在生命科学中的应用展示了广阔的前景。Using a Dual-View wavelength splitter,double-stained African green monkey kidney COS-7 cells,transfected with pEGFP-Myosin 15a and costained with Rhodamine-filopodia were observed based on an ICCD（intensified charge couple device） fluorescence micro-imaging systems.Total internal reflection fluorescence microscopy was used to observe the overexpression of Myosin 15a to the tips of the elongation filopodia.An approach to collecting fluorescence in two channels and avoiding spectra crosstalk was employed to observe Myosin 15a and filopodia distribution in African green monkey kidney COS-7 cells.High sensitivity TIRF technology and two channels imaging method provided a wide application in bio-medical studies.

关 键 词：全内反射荧光(TIRF) 双通道 肌球蛋白 微丝 

分 类 号：TH742[机械工程—光学工程] O439[机械工程—仪器科学与技术]