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首页> 美国卫生研究院文献>other >Plastic Versus Glass Support for an Immunoassay on Metal-Coated Surfaces in Optically Dense Samples Utilizing Directional Surface Plasmon-Coupled Emission
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Plastic Versus Glass Support for an Immunoassay on Metal-Coated Surfaces in Optically Dense Samples Utilizing Directional Surface Plasmon-Coupled Emission

机译：塑料与玻璃支持物进行定向表面等离激元耦合发射的光学致密样品中金属涂层表面的免疫测定

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摘要

We compared plastic (polycarbonate) and high-quality glass support materials for gold-coated slides, when performing a model immunoassay against rabbit IgG using fluorescently labeled (AlexaFluor-647) anti-rabbit IgG, and detecting surface plasmon-coupled emission (SPCE) signals. Both, glass and plastic slides were simultaneously coated with a 48-nm layer of gold and protected with a 10-nm layer of silica. The maximum SPCE signal of AlexaFluor-647 was only two- to three-fold smaller on plastic slides than on glass slides. A small difference in the SPCE angles on glass (θF = 55°) and plastic (θF = 52.5°) slides was observed and can be explained with a slightly smaller refractive index of the plastic. We have not found any difference in the angle distribution (sharpness of the fluorescence signal at optimal SPCE angle) for the plastic slide compared to the glass slide. The kinetics of binding was monitored on the plastic slide as well as on the glass slide. Optically dense samples, a 4% red blood cell suspension and a 15% hemoglobin solution, are causing a reduction in the immunoassay SPCE signal by approximately 15% and three times, respectively, and the percentage of the reduction is the same for plastic and for glass slides. We believe that plastic substrates can be readily used in any SPCE assay, with only marginally lower total signal compared to high-quality glass slides.
机译：当使用荧光标记的（AlexaFluor-647）抗兔IgG对兔IgG进行模型免疫测定并检测表面等离激元耦合发射（SPCE）时，我们比较了用于镀金载玻片的塑料（聚碳酸酯）和高质量玻璃支持材料信号。同时将玻璃和塑料载玻片都涂有48 nm的金层，并用10 nm的二氧化硅层保护。 AlexaFluor-647的最大SPCE信号在塑料载玻片上仅比玻璃载玻片小两到三倍。观察到玻璃（θF= 55°）和塑料（θF= 52.5°）载玻片上的SPCE角有微小差异，可以用塑料的折射率稍小来解释。我们没有发现塑料载玻片与玻璃载玻片的角度分布（在最佳SPCE角度的荧光信号的清晰度）有任何差异。在塑料载玻片和玻璃载玻片上监测结合的动力学。光学密度高的样品，4％的红细胞悬液和15％的血红蛋白溶液分别使免疫分析SPCE信号降低了约15％和3倍，塑料和塑料的降低百分比相同幻灯片。我们相信，塑料基板可轻松用于任何SPCE分析中，与高质量载玻片相比，其总信号仅稍低一些。

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