胞内游离钙浓度测定平台的建立  

THE ESTABLISHMENT OF AN INTRACELLULAR FREE CA^(2+) CONCENTRATION MEASURING SYSTEM

作　　者：雷明[1] 曾晓荣[1] 陈桂兰[1] 刘智飞[1] 杨艳[1] 

机构地区：[1]泸州医学院心肌电生理学研究室,四川泸州646000

出　　处：《泸州医学院学报》2010年第5期477-481,共5页Journal of Luzhou Medical College

基　　金：国家自然科学基金资助项目(No.30670763和No.30870903);四川省教育厅基金资助项目(No.2006ZD020)

摘　　要：目的:建立一种简单有效的细胞内游离钙浓度测定平台。方法:用Fura-2作为荧光指示剂标记细胞内Ca2+,在倒置荧光显微镜下交替检测340nm和380nm激发的荧光强度,计算F340/F380代表的Ca2+相对浓度,再校正为标准浓度。结果:给予细胞刺激(如10mM咖啡因),F340/F380迅速增高,表明细胞内游离钙浓度迅速升高。对于快速变化的胞内Ca2+信号,系统反应灵敏、迅速,能很好地捕捉相关钙浓度变化。结论:由此建立起了一种简单有效的细胞内游离钙浓度测定平台,并应用于实际工作。这一平台在平滑肌细胞及心肌细胞游离钙浓度测定中都得到了充分的应用,平台建设中的第一手资料具有重要参考价值。Objective：To establish an effective system for the measurement of intracellular free Ca2＋ concen-tration.Method： Using Fura-2 as Ca2＋ indicator,the fluorescence excited at 340nm and 380nm was detected un-der inverted fluorescence microscope,F340/F380 ratio was calculated which represents the relative Ca2＋ concen-tration,and then standard Ca2＋ concentration was calibrated.Result： F340/F380 steeply increased after stimulated with 10mM Caffeine,indicating the rapid increase of intracellular free Ca2＋ concentration.This system responds quickly and sensitively to rapid changes of intracellular Ca2＋ concentration.Conclusion： An effective system for the measurement of intracellular free Ca2＋ concentration has been established and applied successfully in the mea-surement of intracellular free Ca2＋ concentration of smooth muscle and myocardial cells.The first-hand data from this system will certainly be of value in the future.

关 键 词：FURA-2 钙离子浓度 咖啡因 

分 类 号：Q433[生物学—生理学]