Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces
Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces
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DOI：
10.1016/j.ab.2004.08.034
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发表时间：
2004-11-15
影响因子：
2.9
通讯作者：
Lakowicz, JR
中科院分区：
文献类型：
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作者：
Matveeva, E;Gryczynski, Z;Lakowicz, JR
我们提出了一个通用免疫分析平台，它在银岛膜(SIF)附近使用增强的全内反射荧光，银岛膜是一种由金属(银)颗粒组成的表面涂层。该平台用于模型免疫分析，其中蛋白质抗原，兔免疫球蛋白G被固定在涂有SIF的玻璃表面。检测到荧光染料标记的抗兔抗体与表面抗原结合的信号；测试了不同颜色的染料。与玻片相比，近距离放置荧光团与表面结合的银纳米结构导致SIF上的信号显著增强(高达40倍)。对于实际的分析开发，使用全内反射激发模式具有显著的优势(相对于经典的正面激发)。有限的消逝波激发体积使背景信号最小化成为可能，并且无需任何洗涤步骤即可使用免疫分析。(C)2004 Elsevier Inc.保留所有权利。
We present a generic immunoassay platform that uses enhanced total internal reflection fluorescence in the proximity of silver island films (SIFs), a surface coating consisting of metal (silver) particles. This platform is used with a model immunoassay where a protein antigen, rabbit immunoglobulin G, was immobilized on the SIF-coated glass surface. The signal from a fluorescent dye-labeled anti-rabbit antibody binding to the surface antigen was detected; different color dyes have been tested. Close placement of the fluorophore to surface-bound silver nanostructures results in dramatic signal enhancement (up to 40-fold) on the SIFs as compared with the glass slides. Use of the total internal reflection mode of excitation has significant advantages (over classic front-face excitation) for practical assay development. The limited evanescent wave excitation volume makes it possible to minimize the background signal and use the immunoassay with no need for any washing steps. (C) 2004 Elsevier Inc. All rights reserved.