利用普通荧光显微镜测定神经细胞内游离钙  

Measurement of Intracellular Calcium Concentration in Nerve Cells Using Conventional Fluorescence Microscope

作　　者：王天佑[1,2,3] 范世福[1,2,3] 张思祥[1,2,3] 孙万新[1,2,3] 张莉[1,2,3] 薛绍白[1,2,3] 

机构地区：[1]北京市神经外科研究所 [2]天津大学精密仪器与光电子工程学院 [3]北京师范大学生物系和国家教委细胞增殖及调控生物学开放实验室

出　　处：《基础医学与临床》1997年第4期75-80,共6页Basic and Clinical Medicine

基　　金：卫生部资助;国家教委细胞增殖及调控生物学开放实验室资助

摘　　要：在普通荧光显微镜及电脑之间安装一支光电倍增管，用来测量细胞荧光强度。用Ｈｏｅｃｈｓｔ３３３４２或碘化丙锭染色，比较小鼠精子与肝细胞ＤＮＡ荧光量，证明此装置测量荧光的线性关系良好。以ｆｌｕｏ－３为钙荧光指示剂，进行了小鼠大脑神经细胞静息［Ｃａ２＋］ｉ定量，并于１小时内追踪高Ｋ＋或谷氨酸引起的［Ｃａ２＋］ｉ信号增高的动态过程。结果表明此装置可用于测量单细胞或细胞团块的［Ｃａ２＋］ｉ。lntracellular free Ca 2+ concentration([Ca 2+ ] i)in nerve cells was measured with a photomultiplier tube connected with a conventional fluorescence microscope and a personal microcomputer. The linearity of the measurement was satisfactory when a comparison was made between the relative DNA fluorescence intensity in mouse spermatazoa and hepatocytes stained with either Hoechst 33342 or propidium iodide. Using fluo 3 as a fluorescent Ca 2+ indicator,resting[Ca 2+ ] i in cultured mouse cerebral cortical cells was quantified.The kinetic changes of [Ca 2+ ] i induced by high K + or glutamate were traced within 1 hr. The results indicated that the assembly could be used in the measurement of [Ca 2+ ] i in either single nerve cells or cell aggregates.

关 键 词：钙 神经细胞 荧光显微镜 

分 类 号：R329－33[医药卫生—人体解剖和组织胚胎学]