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稀土上转换发光材料标记抗体的制备及在免疫组化中的应用

Preparation of Up-conversion Luminescent Material Labeled Antibody and Its Application in Immunohistochemistry

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【作者】 陈颖李菲菲李春光潘庆芝崔满华

【Author】 CHEN Ying1,2,LI Fei-Fei3,LI Chun-Guang3,PAN Qing-Zhi3,CUI Man-Hua1* (1.The Second Hospital of Jilin University,Changchun 130041,China; 2.Changchun Obstetrics and Gynecology Hospital,Changchun 130042,China; 3.State Key Laboratory of Inorganic Synthesis and Preparative Chemistry,Changchun 130012,China)

【机构】 吉林大学第二医院长春市妇产医院吉林大学无机合成与制备化学国家重点实验室

【摘要】 采用微波辐射法合成了具有上转换发光特性的六方相纳米粒子NaGdF4:Yb3+,Er3+(UCNPs),其晶粒大小约为65 nm,且粒子在980 nm的激发光下显示绿光(550 nm).进一步在NaGdF4:Yb3+,Er3+纳米晶的表面包覆了一层二氧化硅层,进行氨基功能化后获得了表面共价结合氨基基团的粒径为70 nm的上转换发光纳米微球NaGdF4:Yb3+,Er3+@SiO2-NH2(UCNPs@SiO2-NH2).通过共价键将UCNPs@SiO2-NH2与多克隆抗体免疫球蛋白联接,将标记后的多克隆抗体应用于传统的免疫组化检测子宫内膜腺细胞中基质金属蛋白酶组织抑制剂-4(TIMP-4)蛋白的表达.结果表明,微波合成的稀土上转换发光纳米材料形貌规则且粒径均一,包覆硅壳后材料具有良好的分散性和水溶性,荧光强度高且稳定,在980 nm激发光下对生物组织无背景荧光,可以很好地检测组织中蛋白质的表达.

【Abstract】 The up-conversion luminescent material for optically written display,NaGdF4: Yb3+,Er3+ up-conversion nanoparticies(UCNPs) were prepared under microwave irradiation.The particle size was about 65 nm and the samples exhibited green(550 nm) fluorescence excited by a 980 nm diode laser.The UCNPs@SiO2-NH2 nanocomposites,the size of which was 70 nm,were prepared and conjugated with polyclonal antibidy.Then,the ability of polyclonal antibody fluorescent probes was detected to recognize the expression of tissue inhibitor of metalloproteinases-4(TIMP-4) in the endometrial gland cells by immunohistochemistry.The results show that the as-prepared UCNPs have uniform shape and size distribution.The UCNPs@SiO2-NH2 nanocomposites offer some distinct advantages,such as good distribution and water-solubility,strong and stable upcoversion fluorscence as well as significant auto-fluorescence from biological tissues resting in low signal to background ratio,and are potentially to apply to detect the expression of protein in tissues at 980 nm excitation.

【基金】 吉林大学无机合成与制备化学国家重点实验室开放课题(批准号:2012-07)资助
  • 【文献出处】 高等学校化学学报 ,Chemical Journal of Chinese Universities , 编辑部邮箱 ,2013年04期
  • 【分类号】O657.1
  • 【被引频次】9
  • 【下载频次】515
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