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

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。

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

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-...