大鼠骨髓嗜多染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 Polychromatic Erythrocytes from Rat Marrow

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155-158,182,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大鼠骨髓嗜多染红细胞微核率自动化检测方法。材料与方法 :以环磷酰胺诱导雄性Wistar大鼠微核形成 ,采用吖啶橙(acridineorange,AO)荧光染色 ,分别用荧光显微镜及单激光流式细胞仪检测大鼠骨髓嗜多染红细胞微核率及正染红细胞微核率。结果 :以前置角散射光(FSC)和DNA荧光(FL1)作图 ,大鼠骨髓细胞被分成明显的五个细胞群———有核细胞、嗜多染红细胞、正染红细胞、含微核的嗜多染红细胞及含微核的正染红细胞 ;经环磷酰胺处理的大鼠骨髓嗜多染红细胞微核率及正染红细胞微核率均呈现良好的剂量_反应关系及时效关系 ;人工显微镜计数结果与流式细胞仪检测结果具有良好的相关性(R=0.949 ,P<0.01) ,二者的曲线拟合方程为 ^Y=1.0967+1.0639X(R=0.949 ,P<0.01)。结论BACKGROUND&ATM: To establish an automatic scoring method based on a single-laser flow cytometer for detection of frequencies of micronucleated polychromatic erythrocytes from rat marrow. MATERIAL AND METHODS: Male Wistar rats were exposed to cyclophosphamide(CP)and their frequencies of micronucleated polychromatic eryˉthrocytes(FMNPCE)as well as frequencies of micronucleated normochromatic erythrocytes(FMNNCE)were deˉtected by manual scoring and a single-laser flow cytometer based on AO staining. RESULTS: Using Forward angle scatter(FSC)and DNA fluorescence(FL1)to protract contour configuration histograms,they showed five distinct cell populations which were total nucleated cells,polychromatic erythrocytes,normochromatic erythrocytes,micronucleated polychromatic erythrocytes and micronucleated normochromatic erythrocytes respectively;both manual and flow cytoˉmetric methods demonstrated positive dose-response trends and time-related increase for fMNPCE and fMNNCE in rat bone marrow;comparison of the results by the two methods showed good correlation between them and curve fit analysis showed the equation was Y ^ =1.0967+1.0639X(R=0.949,P<0.01). CONCLUSION: An automated scoring method based on a single-laser flow cytometer for detection of frequencies of frequencies of micronucleated polychromatic erythrocytes from rat marrow is established.

关 键 词：微核实验 流式细胞仪 自动化检测 大鼠 

分 类 号：R994.6[医药卫生—毒理学]