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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (17): 80-84.doi: 10.11924/j.issn.1000-6850.2013-0945

所属专题： 生物技术； 畜牧兽医

• 水产 渔业 • 上一篇    下一篇

激光扫描共聚焦显微镜检测Fluo-3 AM 负载大鼠心肌微血管内皮细胞最佳浓度和时间的探讨

冯波 董虹 张涛 胡格 房克凤 穆祥   

  • 收稿日期:2013-04-02 修回日期:2013-04-23 出版日期:2013-06-15 发布日期:2013-06-15
  • 基金资助:
    提高微血管自律运动振幅的中药成分抗致病因子伤害的机制研究;白头翁汤治疗细菌内毒素病的机理及有效成分研究;中药提高微血管自律运动振幅的生物学意义研究

Detection of Optimal Concentration and Time of Fluo-3 AM Loading Rat Myocardial Microvascular Endothelial Cells by Laser Scanning Confocal Microscope

  • Received:2013-04-02 Revised:2013-04-23 Online:2013-06-15 Published:2013-06-15

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摘要： 为了探讨钙离子荧光探针Fluo-3 AM 对大鼠心肌微血管内皮细胞(Rat Myocardial Microvascular Endothelial Cells RMMECs)最佳装载浓度和时间。将探针Fluo-3AM分别稀释成1、2、3、4、5 μM/L，每一浓度的探针分别装载细胞5、10、15、20 min，应用激光扫描共聚焦显微镜检测细胞内荧光信号的强度、位置、装载探针后细胞的形态变化来观察装载效果。结果表明：1～3 μM/L的Fluo-3AM负载5~20 min均不出现荧光信号；4 μM/L的Fluo-3AM负载5~10 min时即出现荧光信号；5 μM/L的Fluo-3 AM 孵育细胞15 min后出现清晰且分布均匀的荧光信号。Fluo-3AM对贴壁生长的大鼠心肌微血管内皮细胞负载效果主要受探针浓度和负载时间的影响。

关键词: 大浦乡村世界, 大浦乡村世界

Abstract: In order to explore the optimal concentration and the time of Fluo-3 AM loading adherent Rat Myocardial Microvascular Endothelial Cells (RMMECs). Fluo-3AM were diluted to 1, 2, 3, 4, 5 μM/L and each concentration of the probe was mounted cells for 5, 10, 15, 20 min. Laser Scanning Confocal Microscope (LSCM) was used to detect intracellular fluorescence signal. To observe the loading effect according to the fluorescence intensity, position and changes of cells morphology after loading probe. The results showed that: the fluorescence signal did not appear after loading 5 to 20 min by 1 to 3 μM/L Fluo-3AM, the fluorescence signal began to appear after loading 5 to 10 min by 4μM/L Fluo-3AM, Clear and uniform distributed fluorescence signal could be seen after loading 15 min by 5μM/L Fluo-3AM. The effect of Fluo-3AM loading adherent RMMECs was mainly affected by the concentration of probe and loading time.