ZnSe∶Mn/ZnSe-IgG掺杂量子点探针对人肝癌细胞的免疫荧光成像的影响

摘要

Objective Special immune fluorescent imaging of human hepatocellular carcinoma cell line HCCLM9 AFP antigen with water soluble ZnSe:Mn/ZnSe doped-quantum dots modified by 3-Mercaptopropionic acid.Methods ZnSe:Mn/ZnSe doped-quantum dots were modified with 3-Mercaptopropionic acid to prepare water soluble ZnSe:Mn/ZnSe-IgG probes.The characters of these probes were studied through spectra analyses and transmission electron microscope.These probes were used for special immune fluorescent imaging of human hepatocellular carcinoma cell line HCCLM9 alpha fetal protein (AFP) antigen.Methyl thiazol tetrazolium (MTT) assay were used to compare the cytotoxicity between ZnSe:Mn/ ZnSe-IgG and CdSe/ZnSe-IgG probes.Images were obtained through two-photon confocal laser microscope.Photostability of ZnSe:Mn/ZnSe-IgG probes were studied under continuous illumination by a high intensity laser,comparing with the CdSe/ZnSe and organic fluorescence probes.Results These water soluble ZnSe:Mn/ZnSe-IgG probes have good properties such as broad excite spectra,high fluorescence intensity.They have lower cytotoxicity than CdSe/ZnSe probes.They can specifically and effectively recognize tumor markers.The labeling signals of ZnSe:Mn/ZnSe doped-quantum dots were found to be specific and bright.They exhibited exceptional photostability during continuous illumination for long time by a high intensity laser.Conclusion ZnSe:Mn/ZnSe doped-quantum dots are a kind of ideal fluorescent material with spectral characters and photostability better than conventional organic dye,and they also have better biocompatibility than traditional quantum dots,which can be well used in biomedical imaging area.%目的 利用水溶性ZnSe:Mn/ZnSe掺杂量子点优良的光学特性实现对肝癌细胞株HCCLM9甲胎蛋白(AFP)抗原的特异性免疫荧光标记成像.方法 用巯基丙酸修饰的ZnSe∶Mn/ZnSe掺杂量子点,制备水溶性ZnSe∶Mn/ZnSe-IgG复合物探针.荧光、紫外光光谱分析及透射电镜研究其特性.通过间接免疫荧光法,用水溶性ZnSe∶Mn/ZnSe-IgG复合物探针标记肝癌细胞株HCCLM9 AFP抗原.噻唑蓝(MTT)法检测该复合物探针的细胞毒性,并同传统量子点探针比较.双光子共聚焦激光显微镜观察成像,并进行连续激发实验,同传统量子点探针及有机荧光探针比较光稳定性.结果 该水溶性ZnSe∶Mn/ZnSe-IgG复合物探针具有激发光谱宽、荧光强度高的特点,生物相容性较传统量子点好,无明显细胞毒性.用该探针对肝癌细胞成像清晰,其荧光在共聚焦激光连续激发情况下无明显衰减.结论 ZnSe∶Mn/ZnSe掺杂量子点是一种较理想的荧光材料,比传统量子点毒性更小,并且比传统的有机荧光染料具有更优良的光学特性.