其他(光纤免疫生物传感器) 2011

Tapered plastic optical fiber-based biosensor--tests and application.

Biosensors & bioelectronics Beres C, de Nazaré FV, de Souza NC, Miguel MA, Werneck MM
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组成图示

Tapered plastic optical fiber-based b... 传感器构成示意图

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传感器类型

其他(光纤免疫生物传感器)

检测对象

大肠杆菌(Escherichia coli, E. coli)、酵母(Candida guilliermondii)、羊红细胞(lamb erythrocytes);样品基质:无菌生理盐水悬液(300 µL,10^8 cells/mL)

检测原理

塑料光纤(POF)拉锥区使倏逝场暴露于包层外,与周围介质相互作用。当目标细胞与固定在锥表面的特异性抗体结合时,细胞—抗体复合物改变锥区附近局部折射率。周围介质折射率升高时,倏逝场穿透深度和光耦合状态改变,导致透射光功率下降,输出光功率与周围折射率呈反比。880 nm LED 光经 POF 传输,PIN 光电二极管将透射光转换为光电流,再由 AD8032 跨导放大器、PIC16F876A 微控制器和 LabVIEW 软件读出归一化电压。U形弯曲使高阶模以较小入射角到达芯—包层界面,增加倏逝场穿透深度,从而提高灵敏度。该体系未采用 HCR、RCA 等信号放大,主要依赖免疫捕获和折射率变化。

检测灵敏度

测量范围: 1.33–1.39 RIU(0.06 RIU);灵敏度: 直锥 6.1 mV/10−3 RIU(1.330–1.393),U形锥 12.1 mV/10−3 RIU(1.330–1.393);不确定度: 直锥 12.2 × 10−3 RIU,U形锥 1.42 × 10−3 RIU;10^8 cells/mL 细胞响应: 酵母约50 mV、红细胞约40 mV、大肠杆菌约20 mV

效应效果

该传感器对三种细胞均产生可区分响应:10^8 cells/mL酵母、羊红细胞和大肠杆菌孵育50 min后分别引起约50 mV、40 mV和20 mV变化;SEM证实细胞经抗体结合于锥表面。重复性方面,直锥与U形锥在RI 1.33–1.39间进行10次重复测试,曲线几乎重合;标准偏差分别为74 mV和17 mV,对应RI不确定度12.2×10^-3 RIU和1.42×10^-3 RIU。U形锥灵敏度12.1 mV/10^-3 RIU,优于直锥6.1 mV/10^-3 RIU。作者认为其结构简单、成本低、适合水样中细胞检测,但当前灵敏度尚不足以用于商业生物测量,需进一步改进光电部分。

传感器的构成

  • 基底/换能器:多模阶跃折射率塑料光纤(POF,Mitsubishi Eska GH 4001),PMMA 芯与低折射率氟聚合物包层,拉锥区暴露倏逝场
  • 修饰层:蛋白 A 中间层,用于抗体固定
  • 识别元件:抗大肠杆菌、抗 C. guilliermondii 和抗红细胞抗体(7 mg/mL),固定于锥表面
  • 样品池:玻璃瓶或 1 mL 烧杯,使 U 形锥区浸入含细胞样品
  • 光源:GaAlAs LED(880 nm,SFH485P),向 POF 注入光
  • 光电检测:PIN 光电二极管(SFH217F),将透射光转换为光电流
  • 电子读出:AD8032 跨导放大器、PIC16F876A 微控制器和 LabVIEW 软件,输出归一化电压

中文摘要

细胞检测对临床、食品、水和环境样品微生物分析至关重要,但现有方法耗时。塑料光纤(POF)生物传感器为快速、低成本检测提供可行替代。为研究锥度在微生物检测中的灵敏度,本文考察了锥腰直径等几何参数,因为锥区是该类传感器的关键传感元件。研究采用专门开发的拉锥机制备一系列POF锥传感器,并评估几何尺寸分散,以获得最佳拉锥特性,从而提高传感器响应灵敏度。结果显示,U形(弯曲)构型、锥腰直径0.40–0.50 mm的光纤锥表现最佳。这些光纤锥被用作监测装置的核心部分,并经化学处理作为免疫传感器,用于检测细菌、酵母和红细胞。该工作表明,U形POF锥免疫传感器具有结构简单、易于操作和潜在快速检测细胞的优势,为水样等基质中微生物细胞检测提供了可行方案。

英文摘要

Cells detection is crucial in microbiological analysis of clinical, food, water or environmental samples. However, currently employed methods are time consuming. Plastic optical fiber (POF) biosensors consist in a viable alternative for rapid and inexpensive scheme for detection. In order to study the sensitivity of tapers for microbiological detection, geometric parameters are studied, such as the taper waist diameter since the formation of taper regions are the key sensing element in this particular type of sensors. In this study, a series of POF taper sensors were prepared using a specially developed tapering machine, and the dispersion of geometric dimensions is evaluated, aiming to achieve the best tapering characteristics which will provide a better sensitivity on the sensor response. The fiber tapers that presented the finest results were those constructed in U-shaped (bended) configurations, with taper waist diameters ranging from 0.40 mm up to 0.50 mm. These fiber tapers were used as the main section of the monitoring device, and when chemically treated as immunosensors for the detection of bacteria, yeast and erythrocytes.