标本贮存时间和温度对流式细胞术分析淋巴细胞亚群的影响

Effect of Storage Time and Temperature on Analysis of Lymphocyte Subsets by Flow Cytometry

目的:评估样本贮存时间和温度对流式细胞仪分析外周血淋巴细胞亚群影响。方法:流式细胞术分析采血后不同贮存时间和温度对20例随机抽取患者外周血淋巴细胞亚群百分比和平均荧光强度(MFI)的影响。结果:室温和低温贮存60h内淋巴细胞亚群百分比均无明显改变;室温贮存48h、低温贮存36h淋巴细胞荧光抗体MFI可见明显变化,室温贮存24h、低温贮存12h FSCA MFI可见明显下降;低温贮存下T淋巴细胞百分比和MFI比相同时间室温贮存变化更明显。结论:标本采集后室温贮存36h内进行淋巴细胞亚群检测,检测时适当调高FSC电压或降低FSC阈值,可保障检测结果稳定。

[Objective]To evaluate the influence of sample storage time and temperature on the analysis of peripheral blood lymphocyte subsets by flow cytometry.[Methods]Twenty patients were randomly selected to analyze the effects of different storage time and temperature on the percentage and mean fluorescence intensity( MFI) of lymphocyte subsets by flow cytometry. [Results] There was no significant change in the percentage of lymphocyte subsets within 60 h of room temperature and low temperature storage. There were obvious changes in MFI of lymphocyte after 48 h at room temperature and 36 h at low temperature. FSC MFI stored at room temperature for more than 24 h and low temperature for more than 12 h significantly decreased. Moreover,the percentage and MFI of T lymphocyte stored at low temperature had changed more obviously than those stored at room temperature storage at the same time. [Conclusion]The lymphocyte subsets should be detected within 36 h after the specimen is collected and stored at room temperature. During the detection,the FSC voltage should be properly increased or the FSC threshold should be lowered,and the test results were relatively stable.