流式细胞仪检测白细胞髓过氧化物酶的方法研究  

Detection of cytoplasmic antigen MPO by flow cytometry

作　　者：崇慧峰[1] 丁天凌 吴蓓倩 朱萍 

机构地区：[1]安徽省铜陵市人民医院检验科,244000 [2]上海华山医院血液科,200040

出　　处：《检验医学与临床》2011年第6期657-658,661,共3页Laboratory Medicine and Clinic

摘　　要：目的评价流式细胞仪检测胞浆内抗原髓过氧化物酶(MPO)的方法,探讨实验中各因素对检测结果的影响。方法以HL-60细胞为模型,多参数流式细胞仪分析,比较3种不同破膜剂处理及不同处理时间,标本不同放置时间,不同荧光染料对实验结果的影响。结果比较FACS、Fix&Perm、Fixation&Permeabilization 3种破膜剂,得出FACS破膜剂对细胞的前向散射角/侧向散射角(FSC/SSC)影响最小,相对荧光强度(RFI)明显高于其他两者,效果最佳。3种破膜剂均在破膜时间15 min时可达到最佳效果。标本放置在室温条件下,12 h内应对其进行处理,至24 h细胞已出现部分死亡,将影响实验结果的正确性。PE荧光的MPO在荧光强度上明显优于FITC荧光,因此推荐在条件允许的情况下,尽可能地选用PE荧光的MPO抗体。结论流式细胞仪检测MPO的推荐方法:调整样本细胞数,应用FACS破膜剂15 min,标本必须当天完成检测,尽量采用PE标记的MPO抗体。Objective To explore the examination method of cytoplasmic antigen myeloid peroxidase（MPO） and to study the factors affecting the flow cytometry analysis.Methods With HL-60 cells as model,multiple parameters flow cytometry was used to analyze the effect of three kinds of fixation and permeabilization procedure,different color direct immunofluerescence staining and different sample treatment time on the experimental results.Results FACS permeabilization had the best effect,with the minimal influence of forward scatter/side scatter（FSC/SSC） and highest relative fluorescence intensity（RFI）.15 min of permeabilization was enough.In ordinary temperature,the sample should be treated within 12 h,at the most extending to 24 h.PE stained MPO was better than FITC and the PE stained MPO was suggested.Conclusion The standard method of detecting MPO is suggested as follows：using the FACS permeabilization for 15 min,choosing the PE stained antibodies and sample should be completed the detection on the same day.

关 键 词：髓过氧化物酶 流式细胞术 胞浆内抗原 

分 类 号：R392-33[医药卫生—免疫学]