人外周血网织红细胞微核的流式细胞仪自动化检测  

A Study on Flow Cytometric Automatic Detection of Micronucleated Reticulocytes from Human Peripheral Blood

作　　者：孙立平[1] 李德志[2] 刘志谋[2] 杨录军[1] 刘晋祎[1] 曹佳[1] 

机构地区：[1]第三军医大学卫生毒理教研室,重庆400038 [2]第三军医大学附属新桥医院肿瘤治疗中心,重庆400037

出　　处：《癌变．畸变．突变》2004年第3期159-162,182,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家杰出青年基金资助项目(No.30125037) ;军队杰出青年基金资助项目(No.01J014)

摘　　要：背景与目的 :建立一种基于单激光流式细胞仪的人外周血网织红细胞微核率自动化检测方法。材料与方法 :在本室已建立的大鼠骨髓嗜多染红细胞微核流式细胞仪自动化检测体系基础上 ,使用磁式细胞分选技术 ,富集并分离人外周血中的网织红细胞 ,采用吖啶橙(acridineorange,AO)荧光染色 ,用流式仪检测人外周血中的网织红细胞微核率。结果 :磁式细胞分选技术分离到的CD71阳性红细胞占网织红细胞总量的20 %至90 %,产量在不同个体之间存在显著差异 ,但同一个体其分离效果几乎没有差异 ;网织红细胞经过富集后的人外周血样本被分成明显的五个细胞群———网织红细胞、含微核的网织红细胞、成熟红细胞、含微核的成熟红细胞以及有核细胞 ;7名健康人外周血网织红细胞微核率平均值为2.36‰ ,10名接受累积放疗剂量0、4、10、20、30、40及50Gy后的鼻咽癌患者外周血网织红细胞微核率平均值分别为2.97‰、29.37‰、57.90‰、69.05‰、62.58‰、51.19‰及50.07‰。 结论 :成功建立了基于单激光流式细胞仪的人外周血网织红细胞微核率自动化检测体系。BACKGROUND&AIM: To establish an automatic scoring system based on a single-laser flow cytometer for detection of frequencies of micronucleated reticulocytes from human peripheral blood. MATERIAL AND METHODS: Based on the automated scoring system for detection of frequencies of micronucleated polychromatic erythrocytes from rat marrow set up by our laboratory,magnetic cell separation technique was introduced into automatic micronucleus test this time to enrich reticulocytes from human peripheral blood and acridine orange staining was used to calculate the frequencies of micronucleated reticulocytes from human peripheral blood by a single-laser flow cytometer. REˉSULT: 20%to90%,but no significant difference was seen for the same healthy donor within a relative long time during which several separation tests were performed;five distinct cell populations could be clearly seen which were reticulocytes,micronucleated reticulocytes,residual normochromatic erythrocytes,micronucleated normochromatic eryˉthrocytes and nucleated cells respectively;the baseline of frequency of micronucleated reticulocytes of healthy donors was2.36‰,while corresponding to different cumulative dose of0,4,10,20,30,40or50Gy,the mean frequency of micronucleated reticulocytes of nasopharyngeal cancer patients was2.97‰,29.37‰,57.90‰,69.05‰,62.58‰,51.19‰or50.07‰respectively. CONCLUSION: An automatic scoring system based on a single-laser flow cytometer for detection of the frequencies of micronucleated reticulocytes from human peripheral blood has been established.

关 键 词：微核实验 流式细胞仪 自动化检测 人外周血 

分 类 号：R994.6[医药卫生—毒理学]