两种粒径量子点在人舌鳞癌细胞中双色荧光成像的应用研究

目的 研究粒径655 nm和525nm的两种量子点(quantum dots,QDs)对人舌癌Tca8113细胞内2种热休克蛋白( heat shock protein,HSP)进行特异性荧光标记的成像效果和稳定性,为今后的连续动态观察提供实验依据.方法利用QD655nm和QD525nm,对人舌癌Tca8113细胞内HSP90蛋白和HSP70蛋白进行特异性双重荧光标记,激光连续照射2420391 ms,在共聚焦显微镜下同时观测人舌癌Tca8113细胞内的HSP90蛋白和HSP70蛋白表达及分布,用软件Leica Confocal Software测量量子点QD655nm和QD525nm的荧光信号强度变化.结果 激光共聚焦显微镜下可见人舌癌Tca8113细胞内HSP90蛋白和HSP70蛋白均有明显表达,分别表现为红色和绿色荧光,两种蛋白重叠处呈黄色荧光,在激光连续照射中,两种荧光有所衰减,其中QD655nm的荧光强度值下降相对较快、幅度相对较大.结论 量子点荧光标记技术能同时对人舌鳞癌细胞中HSP90蛋白和HSP70蛋白进行双重标记,而且QD655nm与QD525nm都具有较强的光稳定性,均可用于蛋白的长时间动态监测,其中QD525nm的稳定性更好.

Application of two-color immunofluorescence imaging using quantum dots in human tongue cancer cells Tca8113

Objective To evaluate the application of quantum dots(QDs) fluorescence labeling technique on two kinds of heat shock protein(HSP) in human tongue cancer Tca8113 cells,and to compare the light stability.Methods QD655nm and QD525nm were respectively applied to tag Tca8113 strains with indirect immunofluorescence,and the expression of HSP90 and HSP70 was observed under confocal laser microscopy.The fluorescent signal strength was also compared between those from QD655nm and QD525nm.Results With laser scanning microscope QD655nm quantum dots marked HSP90 expression and QD525nm marked HSP70 expression was clearly seen in Tca8113.After 2 420 391ms’ continuous laser exposure,there was no obvious declining of fluorescent markers.Conclusion QD655nm and QD525nm can simultaneously tag HSP90 and HSP70 in Tca8113 cells,QD525nm has better photostability,and they are all appropriate for studies that require a long dynamic observation of physiological changes in cells.