约氏疟原虫子孢子胞内游离钙的检测方法研究  

Determination of sporozoite of Plasmodium yoelii intracellular free calcium with laser confocal scanning microscopy

作　　者：张锡林[1] 段建华[1] 况明书[1] 黄复生[1] 

机构地区：[1]第三军医大学基础部病原生物学教研室,重庆400038

出　　处：《中国人兽共患病杂志》2002年第1期94-96,共3页Chinese Journal of Zoonoses

摘　　要：目的　研究疟原虫子孢子的胞内Ca2 + 的检测方法。方法　应用共聚焦激光扫描显微镜 (CLSM )观察约氏疟原虫子孢子的胞内游离Ca2 + 分布、变化和检测的实验条件。结果　 3μmol/LFluo - 3/Am同时加入 1μl/ml2 5 %PluronicF - 12 7在37℃恒温下 ,孵育分离的子孢子 6 0min即可达到最佳的负载效果。Fluo - 3/Am浓度的提高和孵育时间延长只能增加背景染色 ,若降低孵育浓度和缩短孵育时间则难以达到最佳的负载效果。静息时子孢子胞内Ca2 + 分布均匀 ,均值为 71 2± 4 8nmol/L。结论　应用Fluo - 3/AM和PluronicF - 12 7结合CLSM技术是研究疟原虫子孢子胞浆内游离Ca2 + 的准确、灵敏的方法之一。Aim To determinate the intracellular free calcium concentration and distribution in the sporozoite of Plasmodium yoelii.Methods Distribution of sporozoite intracellular Ca 2+ was observed with the use of Ca 2+ sensitive dye Fluo-3/AM,Pluronic F-127 and confocal laser scanning microscopy(CLSM) to measure the sporozoite intracellular free calcium.Results The best load condition was that the sporozoites were incubated in 3μmol/ml Fluo-3/AM for 60 minutes and 25% Pluronic F-127 (1μl/ml) which was used to prevent the intracellular free calcium of sporozoite from excretion and entering cell organs of Fluo-3/AM.It was effected fluorescent imaging of sporozoite that the concentration of Fluo-3/Am and incubation time were increased or decreased.The intracellular calcium of resting sporozoite was well distributed and the mean value was 71.2±4.8nmol/L (x±s).Conclusion The one of best methods to study intracellular Ca 2+ in sporozoite of Plasmodium yoelii is labeled by Fluo-3/AM,Pluronic F-127 and detected by CLSM.

关 键 词：约氏疟原虫 子孢子 胞内CA^2+ 共聚焦激扫描显微镜 CLSM 检测 

分 类 号：R382.31[医药卫生—医学寄生虫学]