流式细胞仪检测γ射线照射后小鼠骨髓嗜多染红细胞微核率的研究

Frequency of micronucleated polychromatic erythrocyte in mouse bone marrow after γ-ray radiation as detected with flow cytometry

目的应用流式细胞仪检测γ射线照射后小鼠骨髓嗜多染红细胞微核率,建立本实验室快速检测微核率的自动化实验方法。方法将48只雄性BALB/c小鼠随机分为4个照射剂量组,分别给予60Coγ射线一次性全身照射00、.51、和2Gy,于照射后24、48h取骨髓标本,每个剂量和每个时间点均为6只小鼠。采用流式细胞术检测受照小鼠骨髓嗜多染红细胞微核率,以传统显微镜检微核的方法作为对照。结果流式细胞术及显微镜检方法检测的小鼠骨髓嗜多染红细胞微核率均呈良好的剂量依赖性及时间依赖性,并且两种方法的检测结果呈显著正相关(r=0.987,P<0.01)。流式细胞仪检测速度为人工阅片的40倍以上,且能够避免人工镜检人员阅片的主观性,检测结果更为客观。结论采用流式细胞仪检测小鼠骨髓红细胞微核较传统的人工显微镜检测,更加简便、易行、快速,检测结果准确、可信、客观,适用于大规模核辐射伤员的高通量快速生物剂量诊断。

Objective In order to meet the need of rapid diagnosis of biological dosage of individuals exposed to radiation in a large-scale radiation accident,the rate of micronucleated polychromatic erythrocyte（MPE） in mouse bone marrow after irradiation was detected by flow cytometry（FCM）,so as to develop an automatic method of assessment of the rate of MPE.Methods Forty-eight BALB/c mice were randomly assigned into 4 groups（12 each） and received 60Co γ-ray total body irradiation of 0,0.5,1 and 2Gy,respectively.Bone marrow of animals was collected separately at 24h and 48h after irradiation from 6 mice of each group.The rate of MPE in mouse bone marrow was then detected by FCM.As a control,the MPE were also counted under microscope.Results The rate of MPE as detected by both FCM and microscopy showed a significant time-and dose-dependent manner,and a positive correlation was found between the results of the two methods（r=0.987,P〈0.01）.The detection efficiency of FCM was as fast as 40 times of that by microscopy.By avoiding subjective mistakes by the examiner in microscopic method,FCM may give a more objective result.Conclusion Compared with the microscopic method,FCM is more convenient,easy to handle,rapid,reliable and objective in detecting the rate of MPE,and it is applicable in the rapid diagnosis of biological dosage of individual exposed to radiation in a large-scale radiation accident.