流式细胞仪测定血小板胞浆游离钙浓度  

Determination of the level of cytoplasmic free calcium in human platelets with flow cytometry

作　　者：庄明明[1] 温奕欣 刘少列 洪晓鹏[1] 

机构地区：[1]广东省普宁市华侨医院检验科,广东普宁515300

出　　处：《西安交通大学学报（医学版）》2005年第5期508-510,共3页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的建立流式细胞仪测定血小板胞浆游离钙离子浓度[Ca2+]i的方法。方法用Fluo 3-AM作为钙指示剂,在有或无细胞外钙[Ca2+]o存在的条件下,测定静息和凝血酶激活状态下人血小板胞浆游离钙离子浓度[Ca2+]i的变化。结果Fluo 3-AM标记的血小板的平均荧光强度明显增加。凝血酶使负载Fluo 3-AM标记的血小板胞浆[Ca2+]i明显增加,且显示剂量依赖和对[Ca2+]o的依赖。结论凝血酶引起血小板胞浆[Ca2+]i增加的来源主要是[Ca2+]o,可能也有部分是内钙释放的参与。Objective To set up a method to determine cytoplasmic free calcium [Ca^2＋]i ill human platelets with flow cytometry. Methods As a calcium indicator, Fluo 3-AM was used to determine the changes, induced by thrombin, of the level of platelet plasma free calcium [Ca^2＋]i in presence or absence of outside calcium [Ca^2＋]o, in order to elucidate where the platelet plasma free calcium [Ca^2＋]i mainly come from and it＇s role in the activity of platelets. Results The geometric mean of fluorescence intensity of the platelet, labeled with Fluo 3-AM, was increased obviously. The concentration of platelet plasma free calcium [Ca^2＋]i was increased significantly by thrombin in a dose- and [Ca^2＋]o-dependent manner. Conclusion The increased level of platelet plasma free calcium [Ca^2＋]o, induced by thrombin, mainly comes from [Ca^2＋]o and partly from the released [Ca^2＋]o of intraplatelet probably.

关 键 词：流式细胞仪 血小板 钙 凝血酶 

分 类 号：Q257[生物学—细胞生物学]