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首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >Optical Detection of E. coli Bacteria by Mesoporous Silicon Biosensors
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Optical Detection of E. coli Bacteria by Mesoporous Silicon Biosensors

机译：介孔硅生物传感器对大肠杆菌的光学检测

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A label-free optical biosensor based on a nanostructured porous Si is designed for rapid capture and detection of Escherichia coli K12 bacteria, as a model microorganism. The biosensor relies on direct binding of the target bacteria cells onto its surface, while no pretreatment (e.g. by cell lysis) of the studied sample is required. A mesoporous Si thin film is used as the optical transducer element of the biosensor. Under white light illumination, the porous layer displays well-resolved Fabry-Pérot fringe patterns in its reflectivity spectrum. Applying a fast Fourier transform (FFT) to reflectivity data results in a single peak. Changes in the intensity of the FFT peak are monitored. Thus, target bacteria capture onto the biosensor surface, through antibody-antigen interactions, induces measurable changes in the intensity of the FFT peaks, allowing for a 'real time' observation of bacteria attachment.The mesoporous Si film, fabricated by an electrochemical anodization process, is conjugated with monoclonal antibodies, specific to the target bacteria. The immobilization, immunoactivity and specificity of the antibodies are confirmed by fluorescent labeling experiments. Once the biosensor is exposed to the target bacteria, the cells are directly captured onto the antibody-modified porous Si surface. These specific capturing events result in intensity changes in the thin-film optical interference spectrum of the biosensor. We demonstrate that these biosensors can detect relatively low bacteria concentrations (detection limit of 104 cells/ml) in less than an hour.
机译：基于纳米结构多孔硅的无标记光学生物传感器被设计用于快速捕获和检测作为模型微生物的大肠杆菌K12细菌。生物传感器依赖于目标细菌细胞直接结合在其表面上，而无需对研究样品进行预处理（例如通过细胞裂解）。介孔Si薄膜用作生物传感器的光学换能器元件。在白光照射下，多孔层在其反射率光谱中显示出良好分辨的Fabry-Pérot条纹图案。将快速傅立叶变换（FFT）应用于反射率数据会产生一个峰值。监视FFT峰值强度的变化。因此，目标细菌通过抗体-抗原相互作用捕获到生物传感器表面上，从而引起FFT峰强度的可测量变化，从而可以``实时''观察细菌的附着情况。通过电化学阳极氧化工艺制备的中孔硅膜与特异性针对靶细菌的单克隆抗体缀合。通过荧光标记实验证实了抗体的固定化，免疫活性和特异性。一旦生物传感器暴露于目标细菌，细胞就被直接捕获到抗体修饰的多孔硅表面上。这些特定的捕获事件导致生物传感器的薄膜光学干涉光谱的强度变化。我们证明了这些生物传感器可以在不到一个小时的时间内检测到相对较低的细菌浓度（检测极限为10 4 个细胞/ ml）。

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