摘要
目的:探讨流式细胞术(FCM)检测血小板活化的影响因素。方法:采集6名志愿者静脉血,枸橼酸钠为抗凝剂,室温放置不同时间后进行三色免疫荧光标记,FCM检测血小板早期活化标志物纤维蛋白原受体(Fib-R,即PAC-1)和晚期活化标志物P选择素(CD62P)的变化。结果:血小板活化标志物PAC-1和CD62P随着放置时间的延长而表达增加(P<0.05)。抽血后10min和30minPAC-1和CD62P测定值分别相差2.7%和3.5%。且抽血后在不同的活化水平下重复检测多次,结果重复性好,其变异系数(CV)<5%。结论:在室温,枸橼酸钠抗凝条件下,放置时间对血小板活化标志物PAC-1和CD62P的检测结果有较大影响,宜在30min内对采集血液免疫荧光标记后进行流式细胞仪检测。
AIM ： To investigate the affecting factors of detecting platelet activation by flow cytometry （FCM）. METHODS： Using decoagulant of natrium citricum, anticoagnlated peripheral venous bloods from 6 healthy donors were labeled with the method of three - colour immunofluorescence assay. Platelet activation markers fibrinogen receptor （ Fib - R, PAC - 1 ） and P - selectin （CD62P） were measured. In the same time, the reproducibility of FCM was assessed. RESULTS： The platelet activation markers PAC -1 and CD62P at each time point showed significant difference（P 〈0.05）. The ratio was increased with time extending. The positive ratio of PAC - 1 and CD62P immediately measured （ within 10 min ） was 2. 7% and 3.5% less than those at time point of 30 min. The results were measured several times under different activation levels. The coefficient of variation was less than 5%. CONCLUSION： In room temperature and with decoagulant of natrium citricum, if the measurements of PAC - 1 and CD62P are finished within 30 min after sampling, good reproducibility should be achieved.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第3期510-512,共3页
Chinese Journal of Pathophysiology
基金
广东省科技厅科研基金资助项目(No.53064)
广州市医药卫生科技重点资助项目(No.2004Z006)
