光学法检测血小板体外保存期间跨膜电位的变化

发布时间：2018-11-05 06:59

【摘要】：目的建立使用流式细胞仪检测血小板跨膜电位的方法,观察血小板在体外保存期间跨膜电位变化。方法使用电压敏感染料Di BAC4(3)对20(人)份血小板(20 m L/份)染色后,用流式细胞仪测定血小板在静息状态下和完全去极化时的荧光强度,根据能斯特方程计算血小板跨膜电位,并利用此方法观察血小板在体外保存期间跨膜电位的变化情况。结果 20人份新鲜血小板跨膜电位平均值为(-56±4)m V,CV=5.5%,与膜电位钳法得到的结果(-50-60)m V相似,保存1、3、5 d的血小板跨膜电位分别为(-56±4)、(-54±8)和(-46±6)m V,其中血小板跨膜电位在血小板保存1 d时明显高于5 d(P0.05)。结论所建立起的方法用于测定血小板跨膜电位准确、可靠。
[Abstract]:Objective to establish a flow cytometry method to detect the transmembrane potential of platelets and observe the changes of the transmembrane potential of platelets during in vitro preservation. Methods Di BAC4 (3), a voltage-sensitive dye, was used to stain 20 (human) platelets (20ml / phr). The fluorescence intensity of platelets was measured by flow cytometry at rest and at complete depolarization. The transmembrane potential of platelets was calculated according to Nernst equation, and the change of transmembrane potential of platelets during in vitro preservation was observed by this method. Results the mean transmembrane potential of 20 fresh platelets was (-56 卤4) MV CVV 5.5, which was similar to that obtained by membrane potential clamp method (-50-60) m V), and the transmembrane potential of platelets preserved for 3 days was (-56 卤4), similar to that obtained by membrane potential clamp method (-50-60) m V). In (-54 卤8) and (-46 卤6) m V, the transmembrane potential of platelet was significantly higher than that of 5 days after platelet preservation for 1 day (P0.05). Conclusion the established method is accurate and reliable for the determination of platelet transmembrane potential.
【作者单位】： 华东师范大学生命科学学院;上海市血液中心;
【基金】：上海市卫生与计划生育委员会科研项目(20124240) 上海市公益卫生行业专项(201002005)
【分类号】：R446.11
，

本文编号：2311277