Metal-enhanced fluoroimmunoassay on a silver film by vapor deposition
Metal-enhanced fluoroimmunoassay on a silver film by vapor deposition
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DOI：
10.1021/jp0456842
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发表时间：
2005-04-28
影响因子：
3.3
通讯作者：
Lakowicz, JR
中科院分区：
文献类型：
--
作者：
Zhang, J;Matveeva, E;Lakowicz, JR
我们研究了使用金属增强荧光（MEF）检测气相沉积法生成的银膜的荧光免疫分析。银膜的形态通过膜的厚度来控制。在银膜上涂覆二氧化硅层以保护膜并将荧光团与金属表面分离。通过物理吸附将兔免疫球蛋白 G (IgG) 吸附在二氧化硅上，然后将染料标记的抗兔 IgG 与固定化的兔 IgG 结合。观察到荧光团在银薄膜 (2 nm) 上淬灭，在厚膜 (> 5 nm) 上增强，并在 20 nm 处达到饱和（约增强 10 倍）。 MEF 还取决于二氧化硅的厚度，最大值为 10 nm。在 20 nm 银膜上观察到最低的寿命，这与 MEF 的饱和度一致。这些结果显示了在荧光免疫测定中最大限度增加荧光强度所需的银膜的特性。还使用不同染料标记的抗兔 IgG 研究了 MEF 对发射波长的依赖性。
We studied a fluoroimmunoassay using metal-enhanced fluorescence (MEF) detection on silver film generated by vapor deposition method. The morphology of the silver film was controlled through the thickness of the film. A silica layer was coated on the silver film to protect the film and separate the fluorophore from the metal surface. Rabbit immunoglobulin G (IgG) was adsorbed on the silica by physiosorption and then dye-labeled anti-rabbit IgG was bound to the immobilized rabbit IgG. It was observed that the fluorophore was quenched on a thin silver film (2 nm), enhanced on a thick film (> 5 nm), and reached saturation (ca. 10 times enhancement) at 20 nm. The MEF was also dependent on the thickness of the silica with a maximum at 10 nm. The lowest lifetime was observed on the 20 nm silver film, which was consistent with the saturation of MEF. These results showed the properties of a silver film needed for a maximum increase of fluorescence intensity in a fluoroimmunoassay. Dependence of the MEF on the emission wavelength was also studied using different dye-labeled anti-rabbit IgGs.