利用三通道实时荧光成像方法研究单个活细胞凋亡与胞浆pH值变化的关系  

Study of the Relationship between Apoptosis and Intracellular pH in Single Living Cells Using a Three-Channel Real-Time Fluorescence Imaging Method

作　　者：林丹樱[1] 刘晓晨[1] 马万云[1] 

机构地区：[1]清华大学原子分子纳米科学教育部重点实验室,北京100084

出　　处：《光谱学与光谱分析》2009年第6期1581-1585,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(10874099;90919012);教育部科学技术研究重大项目(306020)资助

摘　　要：利用双波长分光器(Dual-View)和自制滤光片滑块,在基于像增强型电荷耦合器件(ICCD)的实时/快速荧光成像系统基础上,建立了一种基于单个ICCD的三通道实时荧光成像方法,同时发展了相应的图像校准处理方法用于消除光谱串扰的影响,并将该方法应用于单个活细胞的研究。利用Annexin V-FITC和SNARF-1两种荧光探针进行双标记,在单细胞水平上对亚硝基谷胱甘肽(GSNO)诱导小鼠胸腺细胞凋亡与其胞浆pH值变化的关系进行实时研究。结果揭示了GSNO诱导的细胞凋亡与细胞发生自发凋亡时胞浆pH值有不同的变化规律,为这种三通道实时荧光成像方法在生物医学领域的应用展示了广阔的前景。Using a Dual View wavelength splitter, a custom-made filter block and only a single intensified charge-coupled device （ICCD）, a three-channel real-time fluorescence imaging method for single living cell research was established based on an ICCD fast fluorescence micro-imaging system. The relevant calibrating method for images was also developed to eliminate the influence introduced by spectral crosstalk. Double-labeled with two fluorescent indicators, Annexin V-FITC and SNARF-1, mouse thymocyte was treated with S-nitrosoglutathione （GSNO） and the relationship between their apoptosis and intracellular pH changes was studied in real time at the single living cell level. The results not only showed the different regularities of intracellular pH changes between apoptosis induced by GSNO and spontaneous apoptosis, but also provided evidence for the application of the three-channel real-time fluorescence imaging method to bio-medical studies.

关 键 词：三通道 实时荧光成像 活细胞 凋亡 胞浆pH值 

分 类 号：TH74[机械工程—光学工程]