快速制备细胞周期样品方法评估和优化  

Evaluation and Optimization of a Rapid Sample Preparation Method for Cell Cycle Analysis

作　　者：吕红霞[1] 杜立颖[1] LU Hongxia;DU Liying(School of Life Sciences, Peking University, Haidian 100871, China)

机构地区：[1]北京大学生命科学学院,北京海淀区100871

出　　处：《实验科学与技术》2019年第1期32-36,共5页Experiment Science and Technology

摘　　要：该文对快速一步法与乙醇固定法制备流式细胞周期样品进行系统比较,并对快速制备方法进行优化。用碘化丙啶快速染色法制备细胞周期样品,平行比较该方法与乙醇固定方法制备样本的测试结果,评估快速样本制备方法的可靠性和样品放置时间对实验结果的影响。实验结果表明,快速染色法与乙醇固定法相比,细胞周期各时相细胞比例及CV值在统计学上均无显著差异,样品放置6天细胞周期各时相百分含量无显著差异,但CV值和荧光强度有明显改变,在快速染液中添加2%～4%血清,能够有效降低细胞粘度、提高高粘度样本的检测速度。This paper systematically compares the rapid one-step method with the ethanol fixation method for the preparation of flow cytometry samples and optimizes the rapid preparation method.Cell cycle samples were prepared by rapid staining with propidium iodide.The test results of this method and ethanol fixation method were compared in parallel.The reliability of the rapid sample preparation method and the influence of sample placement time on the experimental results were evaluated.The experimental results showed that there was no statistically significant difference in the cell ratio and CV value of the cell cycle between the rapid staining method and the ethanol-fixed staining method.There was no significant difference in the percentage of cell cycle phase with in 6-days staining,but CV value and fluorescence intensity were significantly changed.Adding2%~4% serum to the rapid staining solution can effectively reduce the cell viscosity and improve the detection speed of high viscosity samples.

关 键 词：细胞周期 样品制备 碘化丙啶染色 粘连细胞 流式细胞分析 

分 类 号：TP75[自动化与计算机技术—检测技术与自动化装置]