CD71荧光抗体和PI染色的微核流式细胞术检测研究

目的探索CD71荧光抗体和PI染色的微核流式细胞术自动化检测方法。方法采用-85℃甲醇固定细胞,CD71荧光抗体和PI染色,以伯氏疟原虫感染小鼠血细胞为微核模式生物调校流式细胞仪,建立微核流式细胞术检测方法,并对秋水仙素(COL)诱导的小鼠外周血含微核网织红细胞进行了检测。结果所建立的方法能明确分辨外周血含与不含微核的网织红细胞和成熟红细胞,COL处理小鼠的外周血含微核网织红细胞率呈明显的剂量依赖性升高,流式细胞术与显微镜检测的外周血含微核网织红细胞率有良好相关性(r=098)。结论微核流式细胞术检测方法能准确地检测小鼠外周血含微核网织红细胞。

Automatic analysis of micronuclei by flow cytometry using anti-CD71-FITC and PI staining

OBJECTIVE: To explore flow cytometry (FCM) based method for automatic analysis of micronuclei (MN) in cells staining with anti-CD71-FITC and PI. METHODS: Cell were fixed in -85 degrees C methanol and stained with anti-CD71-FITC and PI, instrument was optimized using malaria-infected erythrocytes as biological standards, a FCM based method for automatic analysis of micronuclei was established. In order to estimate the reliability of the method, the frequencies of micronucleated reticulocytes (MN-RET) in peripheral blood of NIH mice treated with colchicine (COL) was detected using the method. RESULTS: Four cell populations, normochromatic erythrocytes (NCE), reticulocytes (RET), micronucleated erythrocytes (MN-NCE) and MN-RET could be clearly resolved on the FL1-FL2 dot plot using this method. COL induced a dose-dependent increases in the frequency MN-RET in peripheral blood. Data obtained manually and automatically showed a strong linear correlation (r = 0.98) as far as the percentage of MN-RET. CONCLUSIONS: The FCM-based automatic technique using FITC-anti-CD71 and PI staining is a reliable tool for micronucleus analysis.