微梁阵列生化传感器研制及检测应用
发布时间：2018-08-30 10:45
【摘要】：微悬臂梁生化传感技术是一项综合微机电系统、生物化学以及力学等多项学科的新型分析方法。这种传感技术以原子力显微镜技术为基础,通过探测微悬臂梁偏转等信号,对悬臂梁表面上发生的生化反应进行实时探测。该传感技术具有灵敏度高、无须标记、实时原位等优点,在生化分析等领域中具有广泛的研究和应用前景。本文以微悬臂梁传感技术为基础,在已有的多种微悬臂梁传感系统平台的基础上,搭建了能够实时检测包含八根微梁的阵列生化传感系统。该系统利用光纤引导激光束作为微梁阵列探测光源,采用时序点亮照明微梁阵列的方式实现对微梁阵列的探测,有效地消除了悬臂梁信号中由温度漂移、震动等因素带来的噪声,实现了高通量的检测。随后对系统进行了多项性能评估和优化设计,有效地降低了信号噪声、提高了检测灵敏度,提升了系统的宜用性。最后在此传感系统平台上利用适配体成功的实现了对微囊藻毒素LR型的定量检测。本论文的主要工作如下:搭建了以多束光纤引导激光束作为悬臂梁探测光源的微梁阵列传感系统,该系统能够实时检测包含八根悬臂梁的微梁阵列的偏转信息。对系统的整体结构、器件的使用进行了详细介绍;对传感系统的光路进行优化选择,有效地减弱了杂散反射光的影响,使激光对准更加方便;设计了结构、体积合理的反应池,节约了样品、方便了实验操作。设计制作了能够与传感系统搭配使用的微梁阵列芯片。对研制的微梁阵列传感系统的信号噪声进行了分析,对激光扫描周期对信号噪声的影响进行研究,确定每个扫描周期内单个激光工作时长不少于400ms;使用商品化的微梁阵列,利用双材料梁的温升效应,对传感系统响应一致性进行评估,测得的各悬臂梁温度响应灵敏度相对偏差不大于7.8%, 一致性良好。使用免疫抗原抗体检测方法对铜离子和瘦肉精进行定量检测,一致性良好,并能够体现微梁阵列中参考组与实验组的对比意义。对水溶液中的汞离子进行定量检测,在同一浓度下检测结果一致性良好,实验中检测浓度最低为0.1ng/mL。对制作的微梁阵列芯片测试和使用。测得悬臂梁曲率半径约为15mm,能够保证探测实验对使用悬臂梁平直度的要求;制作的微梁阵列在温度响应一致性测试中,各悬臂梁偏转量与温度呈良好的线性关系,拟合系数均大于0.99,而且各悬臂梁的温度响应灵敏度相对偏差不大于5.0%,表现出良好的性能。使用制作的微梁阵列对汞离子进行定量检测并与商品化阵列梁的检测结果进行对比,1ng/mL的汞离子使制作的微梁产生偏转量为21.0nm,0.2ng/mL的汞离子是商品化的微梁产生偏转量为39.0nm,制作的微梁阵列刚度偏大,检测灵敏度较低。利用单梁传感平台和修饰了特异性适配体的微梁对有机磷农药甲拌磷进行检测。利用微梁阵列传感系统和修饰了特异性适配体的微梁阵列对微囊藻毒素LR型进行定量检测,检测浓度范围为1-500ng/mL,理论检测极限为0.6ng/mL,提出的该基于适配体的微梁传感方法能够识别自来水中的微囊藻毒素LR型,并能够区别微囊藻毒素的异构体专门用于微囊藻毒素LR型的检测。
[Abstract]:Micro-cantilever biochemical sensor technology is a new analytical method which integrates micro-electro-mechanical system, biochemistry and mechanics. This sensor technology is based on atomic force microscopy technology. It detects the biochemical reaction on the surface of micro-cantilever by detecting the signal of micro-cantilever deflection and so on. With the advantages of high sensitivity, no labeling, real-time in-situ and so on, it has a wide range of research and application prospects in biochemical analysis and other fields. Based on the micro-cantilever sensing technology, and on the basis of many existing micro-cantilever sensing system platforms, an array biochemical sensor system which can detect eight micro-beams in real time is built. Using fiber-optic guided laser beam as detection light source of micro-beam array, the detection of micro-beam array is realized by lighting the micro-beam array in time sequence. The noise caused by temperature drift, vibration and other factors in the cantilever signal is effectively eliminated, and the high-throughput detection is realized. Finally, the LR-type microcystin was quantitatively detected on the platform of the sensor system. The main work of this paper is as follows: A multi-beam optical fiber guided laser beam was constructed as the light source of the cantilever detection. The system can detect the deflection information of the microbeam array including eight cantilevers in real time. The overall structure and the use of the device are introduced in detail. Optimizing the optical path of the sensor system can effectively reduce the influence of stray reflected light and make the laser alignment more convenient. A microbeam array chip is designed and fabricated, which can be used in conjunction with the sensor system. The signal noise of the microbeam array sensor system is analyzed, the influence of the laser scanning period on the signal noise is studied, and the single laser working in each scanning period is determined. It is not less than 400 ms in length; the temperature response consistency of the sensor system is evaluated by using the temperature rise effect of bimaterial beam with commercialized microbeam array. The relative deviation of the temperature response sensitivity of each cantilever beam is less than 7.8%, and the consistency is good. The results of quantitative detection of mercury ion in aqueous solution are in good agreement at the same concentration. The lowest detection concentration in the experiment is 0.1ng/mL. In the temperature response consistency test, the deflection of each cantilever beam has a good linear relationship with the temperature, and the fitting coefficient is greater than 0.99, and the relative deviation of the temperature response sensitivity of each cantilever beam is less than 5.0%, showing a good performance. The results of quantitative detection of mercury ion by beam array are compared with those of commercial array beam. The deflection of the fabricated microbeam is 21.0 nm due to 1 ng/mL mercury ion. The deflection of the commercial microbeam is 39.0 nm due to 0.2 ng/mL mercury ion. The stiffness of the fabricated microbeam array is high and the detection sensitivity is low. Microbeams modified with specific aptamers were used to detect organophosphorus pesticide phorate. Microbeams with specific aptamers were used to detect microcystin LR. The detection concentration range was 1-500 ng/mL and the theoretical detection limit was 0.6 ng/mL. The sensitive method can identify the LR type of microcystins in tap water and distinguish the isomers of microcystins for the detection of LR type of microcystins.
【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TP212
[Abstract]:Micro-cantilever biochemical sensor technology is a new analytical method which integrates micro-electro-mechanical system, biochemistry and mechanics. This sensor technology is based on atomic force microscopy technology. It detects the biochemical reaction on the surface of micro-cantilever by detecting the signal of micro-cantilever deflection and so on. With the advantages of high sensitivity, no labeling, real-time in-situ and so on, it has a wide range of research and application prospects in biochemical analysis and other fields. Based on the micro-cantilever sensing technology, and on the basis of many existing micro-cantilever sensing system platforms, an array biochemical sensor system which can detect eight micro-beams in real time is built. Using fiber-optic guided laser beam as detection light source of micro-beam array, the detection of micro-beam array is realized by lighting the micro-beam array in time sequence. The noise caused by temperature drift, vibration and other factors in the cantilever signal is effectively eliminated, and the high-throughput detection is realized. Finally, the LR-type microcystin was quantitatively detected on the platform of the sensor system. The main work of this paper is as follows: A multi-beam optical fiber guided laser beam was constructed as the light source of the cantilever detection. The system can detect the deflection information of the microbeam array including eight cantilevers in real time. The overall structure and the use of the device are introduced in detail. Optimizing the optical path of the sensor system can effectively reduce the influence of stray reflected light and make the laser alignment more convenient. A microbeam array chip is designed and fabricated, which can be used in conjunction with the sensor system. The signal noise of the microbeam array sensor system is analyzed, the influence of the laser scanning period on the signal noise is studied, and the single laser working in each scanning period is determined. It is not less than 400 ms in length; the temperature response consistency of the sensor system is evaluated by using the temperature rise effect of bimaterial beam with commercialized microbeam array. The relative deviation of the temperature response sensitivity of each cantilever beam is less than 7.8%, and the consistency is good. The results of quantitative detection of mercury ion in aqueous solution are in good agreement at the same concentration. The lowest detection concentration in the experiment is 0.1ng/mL. In the temperature response consistency test, the deflection of each cantilever beam has a good linear relationship with the temperature, and the fitting coefficient is greater than 0.99, and the relative deviation of the temperature response sensitivity of each cantilever beam is less than 5.0%, showing a good performance. The results of quantitative detection of mercury ion by beam array are compared with those of commercial array beam. The deflection of the fabricated microbeam is 21.0 nm due to 1 ng/mL mercury ion. The deflection of the commercial microbeam is 39.0 nm due to 0.2 ng/mL mercury ion. The stiffness of the fabricated microbeam array is high and the detection sensitivity is low. Microbeams modified with specific aptamers were used to detect organophosphorus pesticide phorate. Microbeams with specific aptamers were used to detect microcystin LR. The detection concentration range was 1-500 ng/mL and the theoretical detection limit was 0.6 ng/mL. The sensitive method can identify the LR type of microcystins in tap water and distinguish the isomers of microcystins for the detection of LR type of microcystins.
【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TP212
【参考文献】
相关期刊论文 前10条
1 邬林;周夏荣;吴尚犬;王萍;张青川;伍小平;;新型微梁阵列生化传感器的研制[J];分析化学;2012年04期
2 魏松红;逄若霖;王，
本文编号：2212835
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