目的:对胞质阻滞法与流式细胞术检测淋巴细胞微核率的一致性进行检验并找出对检测结果影响的主要因素。方法:分别以6个浓度丝裂霉素C或硫酸长春碱处理对数生长期TK6细胞并用流式细胞术检测微核率,以胞质阻滞法为参考;分析设门与不同浓度荧光染料叠氮溴乙锭(EMA)对检测结果的影响。结果:两种微核检测方法测得的TK6细胞微核率在相应剂量组中差异无显著性(P〉0.05)。每组实验结果两种方式间存在显著相关性(Rs=1.0,P〈0.01)。影响微核检测的3个门分别为"Light Scatter","FSC VS SYTOX"与"EMA Negative"。采用300μL 0.1 mg/m L EMA染料可更好地测定淋巴细胞微核。结论 :流式细胞术测定淋巴细胞微核与胞质分裂阻滞法测微核的相关性良好,具有快速,简便的优点,三个门的设门原则和EMA染料的体积和浓度成为影响实验的关键因素。
Objective To investigate the consistence between the flow cytometric lymphocyte micronucleus (MN) and the cytokinesis-block micronucleus test (CBMN), and explore the key factors involved in for flow cytometric lymphocyte micronucleus analysis. Methods The TK6 cells were treated by Mitomycin-C or vinblastine sulfate with 6 different concentrations. CBMN as a reference method, the micronucleus rate was detected by the flow cytometry. The effects of gate setting and different concentrations of fluorescent dye ethidium bromide azide on the MN test were studied. Results There was no significant difference at the rate of TK6 ceils MN s in each dose between these two methods(P 〉 0.05),but the positive correlation was significant (both Rs= 1.0,P 〈 0.01).The gate settings in the flow cytometric MN analysis were Light Scatter, FSC VS SYTOX and EMA Negative. Using 300 ILL 0.1 mg/mL EMA can accurately detect the lymphocyte MN rates. Conclusions There is a good correlation between the flow cytometric lymphocyte MN and CBMN. The flow cytometric MN analysis is quick and convenient. The gate setting and the concentration of EMA were the key factors in this method.