全自动SPR分析仪关键技术研究及应用实验

发布时间：2018-05-04 22:06

本文选题：表面等离子共振 + 生物传感器　；参考：《浙江大学》2014年博士论文

【摘要】：当今环境与食品遭受污染问题日益严重,其中有毒有机污染物在环境中分布广泛、种类繁多、浓度极低,实现快速、灵敏、准确的检测分析面临极大挑战。目前的主要检测方法有化学分析法、生物法和免疫分析法,这些方法均存在不同方面的问题,如仪器昂贵、操作复杂、费时耗力等,无法满足不断增长的大规模快速检测分析的需求。生物传感技术,尤其是基于生物芯片的生物传感技术,其中以表面等离子共振(SPR)传感技术为首要代表,近年来已成为国际相关领域关注的热点,而得到快速发展。并成功应用于环境与食品安全检测、临床诊断、药物筛选等众多领域。本论文通过研究SPR分析仪的关键技术,研制了具有自主知识产权的全自动SPR生化分析仪,开展了几种痕量有机污染物的SPR快速检测实验研究取得了较好的结果。本论文将SPR分析仪研制工作分解成多项相关外围技术和关键技术进行研究和设计,并开展了相关的应用检测实验,完成的主要工作为：1.设计小型和全自动两种微流SIA系统,采用空气实现样品-缓冲液的隔离和保护,并解决了在不同压力下,空气段隔离流路体系的精准注射进样问题,使得进样系统运行稳定,样品-缓冲液的隔离和分离效果好。2.针对SPR传感信号温度敏感特性,研究了温度补偿方法。研究和实验结果表明,基于温度变化量的补偿效果不理想,而结合温度变化率的补偿效果有所改善,但依然难以实用化,因此设计了基于TEC和PID调节的恒温控制系统。3.提出多通道SPR传感器的改进和优化方案。将波长633nm光源改为760nm的LED光源,并加上窄带滤光片；将目前8位AD分辨率的面阵CMOS改成16位AD的CCD；从而有效提高传感器的分辨率和动态范围。4.设计互连通道和可控通道两种集成微流芯片,并开发了外部控制电路,满足SPR生物传感检测芯片表面液体流通通道切换等工作要求。5.提出并研究一种“固定边界质心法”的快速SPR共振角计算方法,用实验数据分析比较了已有“固定基线质心法”、“动态基线质心法”与新方法的性能,结果表明该方法具有最优的综合性能。6.研究并实现了一种实时的传感图平滑降噪算法,运用该算法对多个实验数据进行降噪处理,并对算法性能进行分析评价。对于典型的SPR生物传感图,可将噪声降低至原始水平的3.5%,同时几乎没有信号失真和数据延时,降低了仪器的检出限。7.分别运用自行研制的小型SPR分析仪和全自动SPR分析仪,开展了磺胺甲嗯唑、牛血清蛋白、吡虫啉等多种污染物的检测实验,并与Biacore仪器测试结果比对,检测结果良好。
[Abstract]:Nowadays, the problem of environmental and food pollution is becoming more and more serious, among which toxic organic pollutants are widely distributed in the environment, with a wide variety of types and very low concentrations. It is a great challenge to achieve rapid, sensitive and accurate detection and analysis of toxic organic pollutants. At present, the main detection methods are chemical analysis, biological analysis and immunoassay. These methods all have different problems, such as expensive instrument, complicated operation, time-consuming, etc. Unable to meet the growing demand for large-scale rapid detection analysis. Biosensor technology, especially biosensor based on biochip, with surface plasmon resonance (SPR) sensing technology as the primary representative, has become a hot spot in the field of international concern in recent years, and has been rapidly developed. It has been successfully applied in many fields, such as environmental and food safety detection, clinical diagnosis, drug screening and so on. By studying the key technology of SPR analyzer, an automatic SPR biochemistry analyzer with independent intellectual property rights has been developed in this paper. The SPR rapid detection of several trace organic pollutants has been carried out and good results have been obtained. In this paper, the research and development of SPR analyzer is divided into several related peripheral technologies and key technologies for research and design, and related application testing experiments are carried out. The main work accomplished is: 1: 1. Two microflow SIA systems, small and automatic, were designed. The sample buffer was isolated and protected by air, and the problem of accurate injection of air isolation flow system was solved under different pressure, which made the injection system run stably. The isolating and separating effect of sample buffer solution is good. 2. According to the temperature sensitivity of SPR sensor signal, the temperature compensation method is studied. The results of research and experiment show that the compensation effect based on temperature change is not ideal, but the compensation effect based on temperature change rate is improved, but it is still difficult to be applied. Therefore, a constant temperature control system based on TEC and PID regulation is designed. An improved and optimized scheme of multi-channel SPR sensor is presented. The wavelength 633nm light source is changed into the 760nm LED light source, and the narrow band filter is added. The current 8-bit AD resolution array CMOS is changed to the 16-bit AD LED light source, so that the resolution and dynamic range of the sensor are improved effectively. 4. Two integrated microfluidic chips, interconnect channel and controllable channel, are designed, and an external control circuit is developed to meet the requirements of SPR biosensor detection chip surface fluid flow channel switching. 5. A fast SPR resonance angle calculation method for fixed boundary centroid method is proposed and studied. The performances of the existing "fixed baseline centroid method", "dynamic baseline centroid method" and the new method are analyzed and compared with the experimental data. The results show that the method has the best comprehensive performance. 6. A real-time sensor image smoothing noise reduction algorithm is studied and implemented. The algorithm is used to deal with multiple experimental data, and the performance of the algorithm is analyzed and evaluated. For a typical SPR biosensor, the noise can be reduced to 3. 5% of the original level, and at the same time, there is almost no signal distortion and data delay, which reduces the detection limit of the instrument. The experiments of sulfamethoxazole, bovine serum protein and imidacloprid were carried out by using the self-made SPR analyzer and the automatic SPR analyzer, respectively. The results were compared with those of the Biacore instrument.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：TH83;TP212.9

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