[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.
Yang
Ming-Jie
MJ
Centers for Disease Control and Prevention of Guangdong Province, Guangzhou 510300, China. ymingjie@sohu.com
Zhou
Jian-Chang
JC
Li
Zhi
Z
Yang
Xing-Fen
XF
D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D003078:Colchicine / Q000633:toxicity; D005434:Flow Cytometry / Q000379:methods*; D008288:Malaria / Q000097:blood; D008297:Male; D051379:Mice; D015162:Micronucleus Tests / Q000379:methods*; D010962:Plasmodium berghei; D011897:Random Allocation; D011990:Receptors, Transferrin / Q000276:immunology; D012156:Reticulocytes / Q000648:ultrastructure
D015703:Antigens, CD; C068126:CD71 antigen; D011990:Receptors, Transferrin; D003078:Colchicine