反射式光纤SPR生物传感器的研究及测试实验

发布时间：2018-01-12 08:38

本文关键词：反射式光纤SPR生物传感器的研究及测试实验　出处：《山东大学》2017年硕士论文　论文类型：学位论文

【摘要】：光纤表面等离子体共振(SPR)技术因其免标记、实时检测、灵敏度高等特点被应用到生物检测及环境监测等领域,倾斜光纤光栅(TFBG)由于光栅与纤芯轴向存在一定夹角的特殊结构会产生反向传输的包层模可以激发表面等离子体共振,TFBG与SPR技术相结合在生物传感领域得到广泛应用。本文设计了一种反射式TFBG-SPR生物传感器,既具有性能稳定、耐腐蚀、免标记等特点,还具有制作工艺简单、实现二次耦合及现场检测等优势。首先对SPR光纤传感器、TFBG-SPR传感器及反射式光纤传感器的研究现状进行了总结,介绍了反射式SPR传感器的理论基础,分析了反射式TFBG-SPR传感器的基本原理及传感特性,为传感器的制作提供理论基础。其次,对反射式TFBG-SPR光纤传感器进行结构设计,选择合适的制作工艺完成反射式TFBG-SPR传感器的制备,并对制作完成的传感器进行了氯化镁溶液实验,检测传感器的折射率敏感度为564.1nm/RIU,比普通TFBG的灵敏度提高了 245倍,并将该传感器应用到细菌浓度检测实验中,搭建实验平台并实时监测细菌浓度的变化,为后续的细菌测试及细胞实验奠定了基础。最后,将反射式TFBG-SPR传感器应用到两种生物分子实验研究中,分别为DNA分子实验和蛋白质分子实验。DNA分子选取的是耐甲氧西林金黄色葡萄球菌(MRSA)一段耐药基因,将固定探针增覆在传感器的金膜表面制作成反射式TFBG-SPR的DNA分子探针,再与互补探针发生特异性结合反应,通过观察传感器反射谱谐振峰对应的共振波长偏移情况来实现对反应是否进行及反应速度的判断和分析。反射式TFBG-SPR蛋白质分子实验选取半胱胺盐酸盐作为将抗原固定在金膜表面的中间物质,实现兔IgG分子的固定,制备成蛋白质分子探针,进行杂交实验,对相应抗体山羊抗兔IgG实现特异性识别,杂交实验过程中反射谱的共振波长共偏移了 9.5nm,实验效果良好,验证了两种生物分子实验的可行性。
[Abstract]:Optical fiber surface plasmon resonance (SPR) technology has been applied to biological detection and environmental monitoring because of its characteristics of non-marking, real-time detection and high sensitivity. Tilted fiber grating (TFBG) can excite the surface plasmon resonance (SPR) because of the special structure with a certain angle between the grating and the core. The combination of TFBG and SPR technology has been widely used in the field of biosensors. In this paper, a reflective TFBG-SPR biosensor has been designed, which has the characteristics of stable performance, corrosion resistance, no marking and so on. It also has the advantages of simple fabrication process, secondary coupling and field detection. Firstly, the research status of TFBG-SPR sensor and reflective fiber optic sensor are summarized. This paper introduces the theoretical basis of reflective SPR sensor, analyzes the basic principle and sensing characteristics of reflective TFBG-SPR sensor, and provides a theoretical basis for the fabrication of the sensor. Secondly. The structure of reflective TFBG-SPR optical fiber sensor is designed and the appropriate fabrication process is selected to complete the fabrication of reflective TFBG-SPR sensor. The results show that the sensitivity of the sensor is 564.1 nm / r RIUs, which is 245-fold higher than that of conventional TFBG. The sensor is applied to the bacterial concentration detection experiment. The platform is built and the change of bacterial concentration is monitored in real time, which lays a foundation for the subsequent bacterial testing and cell experiment. Finally. The reflective TFBG-SPR sensor is applied to two kinds of biomolecular experiments. DNA molecular test and protein molecular experiment. The drug resistant gene of methicillin-resistant staphylococcus aureus was selected. The fixed probe was added to the gold film surface of the sensor to form a reflective TFBG-SPR DNA molecular probe and then reacted with the complementary probe specifically. By observing the resonance wavelength offset corresponding to the resonance peak of the reflectance spectrum of the sensor, the determination and analysis of whether or not the reaction was carried out and the reaction rate were realized. Cysteamine salt was selected in the molecular experiment of reflective TFBG-SPR protein. The acid salt is the intermediate substance in which the antigen is immobilized on the surface of the gold film. The rabbit IgG molecule was immobilized, protein molecular probe was prepared, hybridization experiment was carried out, and the specific recognition of goat anti-rabbit IgG was realized. The resonance wavelength of the reflectance spectrum deviated 9.5 nm in the hybridization experiment, which proved the feasibility of the two kinds of biomolecular experiments.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TP212.3

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