新型纳米稀土时间分辨荧光探针的制备与应用
发布时间：2018-11-22 08:53
【摘要】：稀土荧光探针具有荧光寿命长、Stokes位移大、发射峰尖锐等优点,结合时间分辨荧光测定技术可有效去除背景荧光的干扰,在复杂生物及环境样品的测定中具有重要的应用价值。近年,时间分辨荧光生物显微成像技术的快速发展又为稀土荧光生物探针的发展与应用提供了新的机遇。在本博士学位论文的研究中,利用稀土荧光配合物作为发光基团,制备了数种新型的纳米稀土荧光生物探针,并考察了其在时间分辨荧光生化分析中的应用性能。将β-二酮-铕(Ⅲ)荧光配合物2,6-BHHD-Eu3+-BPT与硅烷化试剂IPTES共价偶联,利用反相微乳液聚合法制备了一种新型的可见光激发纳米硅胶-铕荧光微粒。这种纳米微粒呈现规则的球形,具有均一的尺寸(42±3 nm)和较长的荧光寿命(346 μs),其激发波长能够从紫外光区延伸到475 nm。将其用于链霉亲和素标记后,成功实现了复杂水样品中鼠隐孢子虫和小隐孢子虫卵囊的时间分辨荧光免疫成像测定。合成了一种可见光激发的铕(Ⅲ)荧光配合物BHHBB-Eu3+-BPT,并利用去核铁蛋白的解离-重组结构特性制备了一种去核铁蛋白包裹铕(Ⅲ)配合物的荧光蛋白Eu@AFt。该蛋白激发峰波长达420 nm,且具有长的荧光寿命(365μs)和优良的生物亲和性,可直接用于活体细胞的时间分辨荧光成像。将该蛋白用线粒体靶向定位基团SPTPP修饰后制备了一种具有线粒体靶向定位作用的荧光蛋白Eu@AFt-SPTPP,并实现了HeLa细胞中线粒体的共聚焦荧光显微镜靶向定位成像。利用去核铁蛋白的结构特性,通过在该蛋白内包裹铽(Ⅲ)荧光配合物PTTA-Tb3+,并在其表面修饰对NO具有特异性响应的罗丹明衍生物分子,制备了一种基于铽(Ⅲ)配合物-罗丹明荧光共振能量转移机理的对NO具有特异性响应的比率型纳米荧光探针Tb@AFt-Rh。该探针具有荧光寿命长、Stokes位移大、选择性和灵敏度高、生物相容性好及可用于比率型荧光检测等优点,被成功用于HepG2细胞及大型蚤内NO的时间分辨荧光成像测定。将铕(Ⅲ)荧光配合物PTTA-Eu3+与超顺磁性钴铁氧化物纳米粒子及叶酸相结合,制备了一种具有荧光-磁性-癌细胞靶向的多功能纳米粒子PTTA-Eu3+-CoFeO-FA,将其用于癌细胞时间分辨荧光成像及昆明鼠磁共振成像测定的结果表明,这种纳米粒子在癌症的荧光-磁共振双模式成像测定方面具有潜在的应用价值。
[Abstract]:Rare earth fluorescence probe has the advantages of long fluorescence lifetime, large Stokes shift, sharp emission peak, and so on. Combined with time-resolved fluorescence measurement technology, the interference of background fluorescence can be effectively removed, and it has important application value in the determination of complex biological and environmental samples. In recent years, the rapid development of time-resolved fluorescence microimaging technology provides a new opportunity for the development and application of rare earth fluorescent biological probes. In this dissertation, several novel fluorescent biological probes were prepared by using rare earth fluorescent complexes as luminescent groups, and their applications in time-resolved fluorescence biochemistry analysis were investigated. A novel visible light-excited silica gel and europium fluorescent particles were prepared by covalent coupling of 尾 -diketo-europium (鈪，
本文编号：2348762
[Abstract]:Rare earth fluorescence probe has the advantages of long fluorescence lifetime, large Stokes shift, sharp emission peak, and so on. Combined with time-resolved fluorescence measurement technology, the interference of background fluorescence can be effectively removed, and it has important application value in the determination of complex biological and environmental samples. In recent years, the rapid development of time-resolved fluorescence microimaging technology provides a new opportunity for the development and application of rare earth fluorescent biological probes. In this dissertation, several novel fluorescent biological probes were prepared by using rare earth fluorescent complexes as luminescent groups, and their applications in time-resolved fluorescence biochemistry analysis were investigated. A novel visible light-excited silica gel and europium fluorescent particles were prepared by covalent coupling of 尾 -diketo-europium (鈪，
本文编号：2348762
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