摘要
目的建立测定血小板激活时细胞内钙离子变化的方法。方法摸索荧光染料Ｆｌｕｏ－３染色血小板内钙离子的适宜条件，应用流式细胞仪测定标记了Ｆｌｕｏ－３／ＡＭ的血小板激活后细胞内钙离子的变化。结果Ｆｌｕｏ－３标记血小板时，随着染料浓度增加和孵育时间延长，基线荧光强度和钙离子载体Ａ２３１８７刺激后的最大荧光强度均提高，但比值基本一致，不受染料条件的影响。无论二磷酸腺苷（ＡＤＰ）还是凝血酶作诱导剂时，染色和未染色血小板之间的最大聚集率均无差异。ＡＤＰ可引起血小板内钙离子增高，并随着ＡＤＰ浓度增加，钙离子荧光强度增强。结论建立了流式细胞仪测定血小板激活时细胞内钙离子变化的方法。
Objective To measure the intracellular Ca ++ changes in activated single platelets was selected and the instracellular Ca ++ changes in activated fluo 3 loaded into living platelets was selected and the intracellular Ca ++ changes in activated fluo 3 loaded platelets were measured.Results The intensity of both baseline and maximal fluorescence increased with the increasing dye concentrations and incubation time,but the ratio between baseline and maximal fluorescence remained constant.The maximum aggregation rate between dyed and non dyed platelets was not significantly different in use of ADP or thrombin.ADP induced the elevation of platelets instracellular Ca ++ fluorescent intensity and Ca 2+ increased with the increasing ADP concentrations. Conclusion Intracellular Ca 2+ changes in activated single fulo 3 load platelets may be detected with flow cytometry.
关键词
血小板活化
钙离子
测定
Pltelet Platelet activation Calcium
