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-论文.pdf

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1、·2570·2013年12月第30December2013．Vo1．30．No．12·实验研究·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陈良冬卓德强冯茂辉喻学锋李雁【摘要】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．I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

2、疫荧光法，用水溶性ZnSe：Mn／ZnSe—IgG复合物探针标记肝癌细胞株HCCLM9AFP抗原。噻唑蓝(M1rr)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

3、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。【关键词】掺杂量子点；肝癌细胞；免疫荧光标记Immuno-fluorescenceimagingofhepatocellularcarcinomacellswithZnSe：Mn／ZnSe-IgGprobesCHENLiang—dong，ZHUODe—qiang，FENGMao—hui，YUXue_厂eng，LIran．CancerPrevention&TreatmentResearchCenterofWuha

4、nUniversity．CollegeofChemistry&MolecularSciences．WuhanUniver—sity，Wuhan430071，ChinaCorrespondingauthor：LIYah，Email：liyansd@yahoo．COrn【Abstract】0bjectiveSpecialimmunefluorescentimagingofhumanhepatocellularcarcinomacelllineHCCLM9AFPantigenwithwatersoluble

5、ZnSe：Mn／ZnSedoped—quantumdotsmodifiedby3-Mercap—topropionicacid．MethodsZnSe：Mn／ZnSedoped—quantumdotsweremodifiedwith3-MercaptopropionicacidtopreparewatersolubleZnSe：Mn／ZnSe-IgGprobes．Thecharactersoftheseprobeswerestudiedthroughspectraanalysesandtransmis

6、sionelectronmicroscope．TheseprobeswereusedforspecialimmunefluorescentimagingofhumanhepatocellularcarcinomacelllineHCCLM9alphafetalprotein(AFP)anti—gen．Methylthiazoltetrazolium(Mrr)assaywereusedtocomparethecytotoxicitybetweenZnSe：Mn／ZnSe—IgGandCdSe／ZnSe-

7、IgGprobes．Imageswereobtainedthroughtwo—photonconfoeallasermicroscope．PhotostabilityofZnSe：Mn／ZnSe—IgGprobeswerestudiedundercontinuousilluminationbyahighintensitylaser．comparingwiththeCdSe／ZnSeandorganicfluorescenceprobes．ResultsThesewatersolubleZnSe：Mn／

8、ZnSe—IgGprobeshavegoodpropertiessuchasbroadexcitespectra，highfluorescenceintensity．TheyhavelowercytotoxicitythanCdSe／ZnSeprobes．Theycanspecificallyandeffectivelyrecognizetumormarkers．ThelabelingsignalsofZnSe：Mn／ZnSedoped—quantumd