流式细胞术检测细胞存活率的方法学建立  

Development of an assay for the detection of the survival rates of cells by flow cytometry

作　　者：刘阳[1] 赖翼[1] 李敏惠[1] 杨淑霞[1] 郑静[1] 阳泰[1] 李丽梅[1] 邓勇[1] 张强[1] 任荣梅[1] 李钱梅[1] 张帅[1] 邹强[1] 李华[2] 

机构地区：[1]成都医学院科研中心,610083 [2]中国人民解放军成都军区总医院肿瘤科,成都610083

出　　处：《国际检验医学杂志》2011年第15期1663-1664,共2页International Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目(No.81072455);教育部科学技术研究重点项目资助(No.209104);教育部新世纪优秀人才支持计划资助(NCET-09-0888)

摘　　要：目的探讨流式细胞术测定细胞存活率的可行性及准确性，为药物筛选、细胞增殖测定、药物毒性检测等提供新的选择。方法使用流式细胞术、CCK8比色法以及台盼蓝染色排除法测定Jurkat细胞存活率。结果使用流式细胞术，细胞浓度大于10^5／mL时，CV值小于5％；细胞浓度小于10^4／mL时，CV值为5％～10％。使用流式细胞术r^2为0．9935，斜率为0．9994；使用CCK8比色法r^2为0．9912，斜率为1．011；使用台盼蓝染色排除法r^2为0．9698，斜率为1．057。结论流式细胞术进样流速稳定，且与活细胞数量成正比，可用于测定细胞存活率。Objective To investigate the feasibility and accuracy of flow cytometry（FCM） for the detection of the survival rates of cells to provide another choice of drug screen,cell proliferation assay and drug toxicity assay. Methods Flow cytometry,CCK-8 and trypan blue staining technique was performed for the detection of the survival rates of Jurkat cells. Results The coefficient of variation（CV） of FCM was less than 5 %, when the cell concentration exceed 105/mL and was 5 %-10%, when the cell concentration was below 10S/mL. The r^2 and slope of FCM were 0. 993 5 and 0. 999 4 respectively,those of CCK-8 were 0. 991 2 and 1. 011 respectively and those of trypan blue staining technique were 0. 969 8 and 1. 057 respectively. Conclusion The flow rate of sample introduction of FCM could be stable and with direct proportion with the amount of viable cells. FCM could be used for the detection of the survival rate of cells.

关 键 词：流式细胞术 辛卡利特 台盼蓝染色排除法 细胞流速 

分 类 号：R446.6[医药卫生—诊断学]