小鼠骨髓嗜多染红细胞微核单激光流式细胞仪自动化检测  

Flow Cytometer Automatic Methods to Detect Micronucleated Polychromatic Erythrocytes in Mice Bone Marrow

作　　者：胡冰冰[1] 杨录军[1] 周燕虹[1] 刘胜学[1] 刘晋[1] 曹佳[1] 

机构地区：[1]第三军医大学预防医学院卫生毒理学教研室,重庆400038

出　　处：《癌变．畸变．突变》2006年第1期19-22,共4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：背景与目的建立快速检测小鼠骨髓嗜多染红细胞微核的流式细胞仪自动化方法。材料与方法以环磷酰胺处理雄性KM小鼠,分别用丫啶橙(Acridine orange,AO)、单激光流式细胞仪(Flowcytometer,FCM)、姬姆萨(Giemsa)法检测小鼠骨髓及外周血嗜多染、正染红细胞微核率,将检测结果进行比较。结果以前置角散射光(Forward angle scatter,FSC)和DNA荧光(FL1)作图,清晰可见小鼠骨髓细胞被分成5个群体——有核细胞、嗜多染、正染、含微核的嗜多染及正染红细胞;环磷酰胺诱导的小鼠骨髓嗜多染及正染红细胞微核率呈良好的剂量-反应关系;FCM、快速AO法与Giemsa法检测结果具有良好的相关性(r=0.973,P<0.01;r=0.978,P<0.01)。结论AO-FCM自动化检测方法完全适用于小鼠骨髓红细胞微核率的检测。BACKGROUND ＆ ATM： To establish a rapid automatic scoring method for micronucleated polychromatic erythrocytes in mice bone marrow by flow cytometer. MATERIAL AND METHODS： Treating male KM mice with cyclophosphamide（CP） and counting micronucleated cells in bone marrow and peripheral blood either by a single-laser flow cytometer（FCM） acridine orange（AO） or Giemsa, comparing the results from different methods. RESULTS： Contour configuration histograms of FLl（indicating DNA fluorescence） vs. FSC（Forward angle scatter, indicating cell size） were used to show clearly total nucleated cells（TNC）, polychromatic erythrocytes（PCE）, normochromatic erythrocytes（NCE）, micronucleated polychromatic erythrocytes （MNPCE） and micronucleated normochromatic erythrocytes （MNNCE） populations. CP-treated fMNPCE and fMNNCE displayed good close-response relationship. Good correlation was demonstrated between the FCM and manual MNPCE data （r = 0.973,P 〈 0.01;r = 0.978,P 〈 0.01）. C0ONCLUSION： The AO-FCM method developed in our department is applicable to detect micronucleated erythrocytes in mice bore marrow.

关 键 词：丫啶橙 微核试验 流式细胞仪 小鼠 

分 类 号：R994.6[医药卫生—毒理学]