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利用细胞培养技术,Ca2+敏感荧光探针Fluo-3/AM标记细胞,激光扫描共聚焦显微镜检测鸡胚胎巨噬细胞内游离钙浓度(〔Ca2+〕i)的变化。结果显示细菌脂多糖(LPS)可引起培养的鸡胚胎巨噬细胞内〔Ca2+〕i升高,LPS的作用可被维拉帕米部分抑制。本实验表明激光扫描共聚焦显微镜结合Fluo-3/AM能准确迅速地检测单个或多个贴壁粘附细胞胞内〔Ca2+〕i的动态变化,并同步观察细胞的形态及其荧光分布情况。
The changes of intracellular free calcium concentration ([Ca2 +] i) in chicken embryo macrophages were detected by laser scanning confocal microscopy using cell culture technique and Fluo-3 / AM labeled cells with Ca2 + -specific fluorescent probe. The results showed that bacterial lipopolysaccharide (LPS) caused the increase of [Ca2 +] i in cultured chicken embryo macrophages, and the effect of LPS was partially inhibited by verapamil. This experiment shows that laser scanning confocal microscopy combined with Fluo-3 / AM can accurately and rapidly detect intracellular [Ca2 +] i dynamic changes in single or multiple adherent adherent cells, and simultaneously observe the cell morphology and fluorescence distribution.