[使用抗CD71-异硫氰酸荧光素和碘化丙啶染色通过流式细胞术自动分析微核]

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

作者信息

Yang Ming-Jie, Zhou Jian-Chang, Li Zhi, Yang Xing-Fen

机构信息

Centers for Disease Control and Prevention of Guangdong Province, Guangzhou 510300, China.

出版信息

Wei Sheng Yan Jiu. 2005 Jan;34(1):25-8.

PMID:15862014

Abstract

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.

摘要

目的

探索基于流式细胞术（FCM）的方法，用于自动分析抗CD71 - FITC和PI染色细胞中的微核（MN）。

方法

细胞用 -85℃甲醇固定，并用抗CD71 - FITC和PI染色，以感染疟原虫的红细胞作为生物学标准对仪器进行优化，建立基于FCM的微核自动分析方法。为评估该方法的可靠性，用该方法检测秋水仙碱（COL）处理的NIH小鼠外周血中含微核网织红细胞（MN - RET）的频率。

结果

使用该方法在FL1 - FL2散点图上可清晰分辨出四个细胞群体，即正色红细胞（NCE）、网织红细胞（RET）、含微核红细胞（MN - NCE）和MN - RET。COL诱导外周血中MN - RET频率呈剂量依赖性增加。就MN - RET百分比而言，手动和自动获得的数据显示出很强的线性相关性（r = 0.98）。