激光扫描共聚焦显微镜术检测单个血小板钙波动方法的探讨

目的 探讨血小板贴壁的简易方法，同时，观察单个血小板激活前后细胞内钙波动及形态变化的规律。方法 采用多种粘附剂固定血小板，利用钙荧光探针（Fluo-3/AM)（终浓度10－20μmol)进行染色，在激光扫描共聚焦显同镜检测下加入二磷酸腺苷，观察和分析血小板内Ca^2 浓度及形态变化。结果 多聚赖氨酸促进血小板贴壁固定的效果最佳；单个血小板活后细胞内Ca^2 浓度为激活前的128％，形态圆形或椭形转为不规则形，有空泡，突起形成。结论 多聚赖氨酸是血小板贴壁的理想粘附剂，本方法能简易，快捷地监测血小板激活过程中胞浆内钙离子动态变化及形态改变，有助于血小板功能的深入研究。

The Method of Platelets Strapping and Platelets Intracellular Calcium Detection

Objects:To explore the simple methods of platelets strapping and observe the platelets intracellular calcium oscillations before and after the platelets be simulated.Methods:Adopt a few adhesives fixed platelets,stained with Fluo-3/Am(10～20umol/L),detect the change of Ca 2+ with Laser Scanning Confocal Microscopy(LSCM).Results:Polylysine is the best adhesive,it can detect the change of Ca 2+ directly using LSCM,Ca 2+ oscillations,is about 128% after the platelets be simulated,its morphologic change from round,elliptic to irregular with vacuolar synaptic shape.Conclusions:Polylysine is a better adhesive for platelets strapping.This method is simple,fast and effective method of detecting Ca 2+ oscillations in simulated platelets,beneficiated to platelets functional research.