摘要
目的利用巯基乙酸修饰的水溶性量子点(QDs)优良的光学特性实现对肝癌细胞株HCCLM6 AFP抗原的特异性免疫荧光成像。方法用巯基乙酸修饰的QDs,制备水溶性QDs-IgG复合物探针。荧光、紫外光光谱分析、琼脂糖凝胶电泳及透射电镜研究其特性。通过间接免疫荧光法,用水溶性QDs-IgG复合物探针特异性识别肝癌细胞株HCCLM6 AFP抗原。将体外培养的肝癌细胞株HCCLM6皮下接种BALB／C-nu／nu裸小鼠。手术取瘤,石蜡切片,水溶性QDs-IgG复合物探针免疫荧光成像。双光子共聚焦激光扫描荧光显微镜观察成像,并与FITC荧光进行连续激发实验比较光稳定性。结果该水溶性QDs-IgG复合物探针具有激发光谱宽、荧光强度高的特点,能与特异性抗原良好结合。用该探针对肝癌细胞及组织成像清晰,其荧光在共聚焦激光连续激发1 h情况下无明显衰减,而FITC荧光在1 h内基本猝灭。结论QDs比传统的有机荧光染料具有更优良的光学特性,适用于生物医学成像领域。
Objective Special immune fluorescent imaging of human hepatocellular carcinoma cell line HCCLM6 AFP antigen with water soluble quantum dots （QDs） modified by thioglycolic acid. Methods QDs were modified with thioglycolic acid to prepare water soluble QDs-lgG probes. The properties of these probes were studied through spectra analyses, agarose gel electrophoresis and transmission electron microscopy. These probes were used for special immune fluorescent imaging of human hepatocellular carcinoma cell line HCCLM6 AFP antigen. HCCLM6 cells were inoculated subcutaneously into nude mice. Tumors were taken out and embedded in paraffin for section and immuno-fluorescence imaging. Images were obtained through two-photon confocal laser scanning fluorescence microscopy. Photostability of QDs was compared with FITC after continuous illumination for 1 h by a high intensity laser. Results The water soluble QDs-IgG probes had good properties, including broad excite spectra, high fluorescence intensity. They could specifically and effectively recognize tumor markers. The labeling signals of QDs were found to be more specific and brighter than those of FITC. They exhibited extraordinary photostability during continuous illumination for 1 h by a high intensity laser, while the FITC signals faded very quickly to undetectable. Conclusion QDs are ideal fluorescent materials with spectral properties and photostability better than conventional organic fluorescent, making them very suitable for biomedical imagingarea.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第9期1085-1087,i0004-i0005,共5页
Chinese Journal of Experimental Surgery
基金
教育部新世纪优秀人才支持计划资助项目(NCET-04-0669)
全国优秀博士学位论文作者专项资金资助项目(200464)
关键词
量子点
癌
肝细胞
免疫荧光
Quantum dots
Carcinoma, hepatocellular
Immuno-fluorescence
