NK细胞胞浆内游离Ca^(2+)浓度和分布的测定  

DETERMINATION OF NK INTRACELLULAR FREE CALCIUM CONCENTRATION AND DISTRIBUTION

作　　者：邬于川[1] 张锡林[2] 李程旭[1] 

机构地区：[1]泸州医学院微生物学与免疫学教研室,泸州646000 [2]第三军医大学病原生物学教研室

出　　处：《泸州医学院学报》2001年第3期183-186,共4页Journal of Luzhou Medical College

基　　金：四川省教委重点课题!A96 0 2 43

摘　　要：目的 :确定NK细胞胞浆内Ca2 +的浓度及分布的检测方法。方法 :应用共聚焦激光扫描显微镜(CLSM)观察人外周血中NK细胞胞浆内Ca2 +的分布 ,并用Fluo- 3/AM和PluronicF - 12 7对胞浆内游离Ca2 +的浓度进行测定。结果 :最佳负载条件是用 2 μmol/ml的Fluo - 3/AM在 37℃恒温下孵育 6 0分钟 ,同时加入 1μl/ml 2 5 %的PluronicF - 12 7以阻断Ca2 +荧光示踪剂Fluo - 3/AM的外排和房室化。静息状态下NK细胞胞浆内Ca2 +分布不均衡 ,Ca2 +浓度为 91 3± 4 8nM( x±s)。结论 :应用Fluo - 3/AM和PluronicF - 12 7结合CLSM技术是研究NK细胞胞浆内游离Ca2 +的准确 ,简单 ,灵敏的方法之一。Objective: To determine the intracellular free calcium concentration and distribution in human NK cells. Methods: Distribution and concentration of human NK intracellular Ca 2+ were observed and determined by means of using Ca 2+ sensitive dye Fluo-3/AM, Pluronic F-127 and confocal laser scanning microscopy(CLSM). Results: The best load condition was that the NK cells were incubated in 2μmol/ml Fluo-3/AM for 60 minutes, and 25% Pluronic F-127(1μl/ml) was used to prevent Fluo-3/AM from excretion and compartmentation. It was found that the intracellular calcium distribution in resting NK cells was not even and NK [Ca 2+ ]i was 91.3±4.8n M(±S). Conclusion: The use of Fluo-3/AM, Pluronic F-127 and CLSM is one of the best methods to study intracellular Ca 2+ of NK cells.

关 键 词：NK细胞 CA^2+ 共聚焦激光扫描显微镜 胞浆内 浓度 分布 测定 

分 类 号：R392.12[医药卫生—免疫学] R329.2[医药卫生—基础医学]