摘要
为了探讨Ca2+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Fluo-3 AM分别稀释成1、2、3、4、5μM/L,每一浓度的探针分别装载细胞5、10、15、20min,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来观察装载效果。结果表明:1～3μM/L的Fluo-3 AM负载5～20min均不出现荧光信号;4μM/L的Fluo-3 AM负载5～10min时即出现荧光信号;5μM/L的Fluo-3 AM孵育细胞15min后出现清晰且分布均匀的荧光信号。Fluo-3 AM对贴壁生长的RMMECs负载效果主要受探针浓度和负载时间的影响。
In order to explore the optimal Microvascular Endothelial Cells （RMME concentration of the probe was mounted concentration and the time of Fluo-3 AM loading Rat Myocardial Cs）. Fluo-3AM were diluted to 1, 2, 3, 4, 5 μM/L and each 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 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 rain by 4 μM/L Fluo-3AM, Clear and uniform distributed fluorescence signal could be seen after loading 15 rain by 5 μM/L Fluo-3AM. The effect of Fluo-3AM loading adherent RMMECs was mainly affected by the concentration of probe and loading time.
出处
《中国农学通报》
CSCD
2013年第17期80-84,共5页
Chinese Agricultural Science Bulletin
基金
北京市教委科技发展计划重点项目"中药提高微血管自律运动振幅的生物学意义研究"(KZ201110020021)
国家自然科学基金"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"(31101850)
北京市教委人才强教深化计划-学术创新团队"白头翁汤治疗细菌内毒素的机理及有效成分研究"(PHR201007142)
科研基地-科技创新平台"抗菌中兽药成分组方原则及新兽药研发"(PXM2012_014207_000013)
科研基地-科技创新平台"磷酸二酯酶4抑制剂对肺微血管内皮细胞黏附分子表达研究"(PXM2012_014207_000010)
