文章摘要
李向阳,华慧,颜超,于倩,郑葵阳,汤仁仙.流式细胞术检测血清细胞因子常见问题与对策[J].济宁医学院学报,2018,41(1):73-76
流式细胞术检测血清细胞因子常见问题与对策
Problems and strategies in cytokine testing with flow cytometry
投稿时间:2017-12-19  
DOI:10.3969/j.issn.1000-9760.2018.01.018
中文关键词: 流式细胞术;CBA;细胞因子
英文关键词: Flow Cytometry;CBA;Cytokine
基金项目:国家自然科学基金项目(81171590,31302077);徐州医科大学校级课题(2014KJ09)
作者单位E-mail
李向阳 徐州医科大学基础医学院, 徐州 221004  
华慧 徐州医科大学基础医学院, 徐州 221004  
颜超 徐州医科大学基础医学院, 徐州 221004  
于倩 徐州医科大学基础医学院, 徐州 221004  
郑葵阳 徐州医科大学基础医学院, 徐州 221004  
汤仁仙 徐州医科大学基础医学院, 徐州 221004 tangrenxian-t@163.com 
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中文摘要:
      目的 分析流式CBA法检测细胞因子常见问题及处理对策。方法 运用流式微球捕获芯片技术(cytometrin bead array,CBA)法对华支睾吸虫感染小鼠血清中Th1/Th2/Th17相关细胞因子进行检测,分析常见问题,探讨相关对策。结果 采用相关实验方法,上调FSC/SSC的阈值,可以有效减少细胞碎片的干扰,降低噪音;在样本处理时,加强对微球的涡旋混匀,避免了微球聚集现象,同时有利于样本与微球更好的结合;通过增加标准品的稀释点数,扩大了检测范围,保证了对低浓度样本的检测;通过对目标微球圈门设定,并导出相应数据,FCAP软件能够对实验数据进行有效的分析,得到较好的实验结果,提高实验的准确性和可行性。结论 建立了流式CBA法检测血清细胞因子常见问题的处理方法及对策。
英文摘要:
      Objective To analyse common problems and countermeasures in the detection of cytokines by CBA.Methods The levels of Th1/Th2/Th17 cytokines of mice serum affected by clonorchis sinensis were detected with Th1/Th2/Th17 Cytokine Kit.Then the common problems were analysed and the relevant countermeasures were explored.Results By using the correlation processing method,the increasing the FSC/SSC threshold reduced the interference of cell debris effectively.During the sample processing,the beads were mixed thoroughly by vortex,which could avoid the the phenomenon of aggregation and be in favor of the combination of beads and sample.By increasing the number of standard levels,the scope of testing was expanded and the low concentration samples could be detected.By exporting target bead population the experimental data could be analyzed by FCAP software effectively,and the experimental results were better,which could improve the accuracy and feasibility of the experiment.Conclusion The method and countermeasure of the common problems in the detection of serum cytokines by flow CBA assays were established.
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