活细胞钙离子浓度荧光显微检测系统的研制和应用  

Development and Application of the Fluorescent Microscopy System for Detecting [Ca ^(2+)]_i in Living Cells

作　　者：兰李[1] 周云燕[1] 杨志勇[1] 瞿安连[1] 徐涛[1] 

机构地区：[1]华中科技大学生命科学与技术学院生物物理与化学研究所,湖北武汉430074

出　　处：《生物医学工程研究》2004年第2期114-117,共4页Journal Of Biomedical Engineering Research

基　　金：国家自然科学基金资助项目 (30 32 70 0 1);国家自然科学基金资助重点项目 (30 130 2 30 )

摘　　要：研制了活细胞钙离子浓度荧光显微检测系统。该系统的工作原理是 :氙灯发射的光线被椭球面镜聚焦后以衍射光栅分光 ,通过光纤、双路耦合聚光镜和物镜将单色光纤入样本平面以激发细胞中的钙离子荧光探针发出荧光 ,最后用光电倍增管检测荧光信号。使用该系统 ,测定了高K+ 去极化刺激下PC12细胞 (大鼠嗜铬细胞瘤细胞 )内 [Ca2 + ]i 变化的动态特性曲线。结果表明 :静息状态下PC12细胞内 [Ca2 + ]i 为 80 .0±15 .0nmol/L ,高K+ 去极化刺激可使PC12细胞内 [Ca2 + ]i 增加到 1.9± 0 .2 μmol/L ,上升至峰值时间约为2 0 .1s。表明此系统可以满足检测活体细胞 [Ca2 + ]i 的需要。The fluorescent microscopy system to detect [Ca 2+] i in living cells was build. In this system, the light from the xenon light source was converged by elliptical mirror and diffracted by diffraction grating; then the optical fiber and dual_coupling condensor transmitted monochromatic light to the specimen plane to generate the fluorescence of the fluorescent probe within the cell; at last the fluorescent signal was detected by the photomultiplier tube. Using this system, the dynamic curve of [Ca 2+] i in PC12 cells after high K+ depolarization was given. The results showed that the resting levels of [Ca 2+] i in PC12 cells were 80.0±15.0nmol/L. After high K+ depolarization was given, the concentrations of intracellular free [Ca 2+] i in PC12 cells were raised by 1.9±0.2μmol/L, and the time of high-K+ depolarization stimulating [Ca 2+] i elevation to a peak point was about 20.1s. These results verify that this system is suitable for detecting [Ca 2+] i in living cells.

关 键 词：[CA^2+]I 显微 PC12细胞 钙离子浓度 刺激 瘤细胞 嗜铬细胞瘤 活细胞 活体细胞 荧光 

分 类 号：Q945[生物学—植物学] R446[医药卫生—诊断学]