基于微图案硅基晶片的生物检测研究
发布时间：2019-03-13 08:46
【摘要】：生物芯片和微阵列是生物检测技术的一个重要发展趋势。在芯片制备中，选择合适的基片材料与点阵排列，降低点阵与背景、点阵与点阵之间的信号干扰，是需要解决的课题之一。本文设计了微图案SiO_2/Si硅基晶片和微图案Au/SiO_2硅基晶片两种基材，对其表面处理、生物检测可行性以及在相关光学检测方法中的应用进行了探讨，主要研究内容如下： 1、微图案SiO_2/Si硅基晶片的表面处理与修饰，采用浓H_2SO_4/H_2O_2、浓HCL/H_2O_2/H_2O、NH_4OH/H_2O_2/H_2O三种清洗液对其表面进行亲水化处理，结果显示H_2SO_4/H_2O_2溶液的处理效果最好，最佳处理时间为30分钟。同时对微图案SiO_2/Si晶片表面的共价修饰方法和疏水化处理方法进行了比较，结果表明，前者对蛋白质的固定效果优于后者。对微图案Au/SiO_2晶片，采用MUA修饰其表面，接触角测量结果表明，Au/SiO_2晶片表面已成功修饰MUA，可进一步与生物分子连接。 2．微图案SiO_2/Si晶片和微图案Au/SiO_2晶片用于蛋白质荧光检测。结果表明，两种晶片固定人IgG底物的最佳浓度均为100mg/L，两种晶片均在图案区获得相当强的荧光信号，而非图案区背景信号很低，可作为基材用于蛋白质芯片检测。对两种硅基晶片进行表面元素的XPS分析，证明蛋白质已与晶片图案区很好地结合。对蛋白质的检测结果表明，微图案SiO_2/Si硅基晶片和Au/SiO_2硅基晶片表面的蛋白质检测灵敏度均为10μg/L。 3．微图案SiO_2/Si晶片和微图案Au/SiO_2晶片用于DNA荧光检测。两种微图案硅基晶片均在图案区获得相当强的荧光信号，而非图案区背景信号很低，表明SiO_2/Si晶片和Au/SiO_2晶片用于DNA检测，可完全排除背景信号的干扰，确定了SiO_2/Si晶片和Au/SiO_2晶片可作为其它DNA检测方法的芯片基材而进一步推广应用。分别考察了两种晶片的DNA检测灵敏度，结果表明，微图案SiO_2/Si晶片表面的DNA检测限为10pM，Au/SiO_2晶片表面的DNA检测限为100pM。 4、采用椭偏检测和双光束干涉光学检测法对微图案SiO_2/Si硅基晶片表面蛋白质膜层厚度进行分析。椭偏检测的结果显示，SiO_2/Si硅基晶片表面蛋白质膜层的平均厚度为4.3nm；通过双光束干涉光学检测系统检测不同浓度的兔抗人IgG所输出的信号，，结果显示其厚度增加约4.27nm，与椭偏检测的结果相吻合。这表明微图案SiO_2/Si硅基晶片可以应用于免标记的光学检测系统。
[Abstract]:Biochip and microarray are an important development trend of biological detection technology. In the chip fabrication, choosing the appropriate substrate material and lattice arrangement, reducing the signal interference between the dot matrix and the background, and the dot matrix is one of the issues that need to be solved. In this paper, two kinds of micro-patterned SiO_2/Si silicon-based wafer and micro-patterned Au/SiO_2 silicon-based wafer are designed, and their surface treatment, biological detection feasibility and application in related optical detection methods are discussed. The main research contents are as follows: 1. The surface treatment and modification of micro-patterned SiO_2/Si silicon wafer were carried out by using concentrated H _ 2O _ 2 so _ 4 / H _ 2O _ 2O _ 2 and HCL / H _ 2O _ 2 O _ 2 O _ (2). The surface of NH_4OH/H_2O_2/H_2O was treated by hydrophilic treatment with three kinds of cleaning solution. The results showed that the treatment effect of H_2SO_4/H_2O_2 solution was the best, and the best treatment time was 30 minutes. At the same time, the covalent modification method and hydrophobic treatment method on the surface of micropatterned SiO_2/Si wafer were compared. The results showed that the former was better than the latter in immobilization of protein. MUA was used to modify the surface of micropatterned Au/SiO_2 wafer. The results of contact angle measurement showed that the surface of Au/SiO_2 wafer modified MUA, could be further connected with biomolecules. 2. Micropatterned SiO_2/Si chips and micropatterned Au/SiO_2 chips were used for protein fluorescence detection. The results showed that the optimal concentration of human IgG substrate was 100 mg / L, the fluorescence signal of both chips was very strong in the pattern region, but the background signal in the non-pattern region was very low, so they could be used as substrate for protein chip detection. The XPS analysis of the surface elements of the two silicon-based wafers shows that the protein binds well to the pattern region of the wafer. The results of the protein test showed that, The sensitivity of protein detection on the surface of micropatterned SiO_2/Si silicon-based wafers and Au/SiO_2 silicon-based wafers is 10 渭 g / L. 3. The micropatterned SiO_2/Si wafers and micropatterned Au/SiO_2 wafers are used for DNA fluorescence detection. Both micropatterned silicon wafers have very strong fluorescence signals in the pattern area, but the background signals in the non-pattern region are very low, indicating that the SiO_2/Si chip and the Au/SiO_2 chip can completely eliminate the interference of the background signal when used for DNA detection. The SiO_2/Si chip and Au/SiO_2 chip can be used as the substrate for other DNA detection methods. The sensitivity of two kinds of wafers for DNA detection was investigated. The results showed that the detection limit of DNA on the surface of the micropatterned SiO_2/Si wafer was 10 pm, and the detection limit of DNA on the surface of the au-Si Sio _ 2 wafer was 100 pm 路cm ~ (- 1). 4. The thickness of protein film on micro-patterned SiO_2/Si silicon wafer was analyzed by ellipsometry and two-beam interferometric optical detection. The results of ellipsometry show that the average thickness of protein film on the surface of SiO_2/Si silicon wafer is 4.3 nm. The two-beam interferometric optical detection system was used to detect the signal output of rabbit anti-human IgG with different concentrations. The results showed that the thickness of the signal was increased by about 4.27 nm, which was consistent with the ellipsometry. This indicates that the micro-patterned SiO_2/Si silicon wafer can be used in the label-free optical detection system.
【学位授予单位】：湖南工业大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.0
本文编号：2439248
[Abstract]:Biochip and microarray are an important development trend of biological detection technology. In the chip fabrication, choosing the appropriate substrate material and lattice arrangement, reducing the signal interference between the dot matrix and the background, and the dot matrix is one of the issues that need to be solved. In this paper, two kinds of micro-patterned SiO_2/Si silicon-based wafer and micro-patterned Au/SiO_2 silicon-based wafer are designed, and their surface treatment, biological detection feasibility and application in related optical detection methods are discussed. The main research contents are as follows: 1. The surface treatment and modification of micro-patterned SiO_2/Si silicon wafer were carried out by using concentrated H _ 2O _ 2 so _ 4 / H _ 2O _ 2O _ 2 and HCL / H _ 2O _ 2 O _ 2 O _ (2). The surface of NH_4OH/H_2O_2/H_2O was treated by hydrophilic treatment with three kinds of cleaning solution. The results showed that the treatment effect of H_2SO_4/H_2O_2 solution was the best, and the best treatment time was 30 minutes. At the same time, the covalent modification method and hydrophobic treatment method on the surface of micropatterned SiO_2/Si wafer were compared. The results showed that the former was better than the latter in immobilization of protein. MUA was used to modify the surface of micropatterned Au/SiO_2 wafer. The results of contact angle measurement showed that the surface of Au/SiO_2 wafer modified MUA, could be further connected with biomolecules. 2. Micropatterned SiO_2/Si chips and micropatterned Au/SiO_2 chips were used for protein fluorescence detection. The results showed that the optimal concentration of human IgG substrate was 100 mg / L, the fluorescence signal of both chips was very strong in the pattern region, but the background signal in the non-pattern region was very low, so they could be used as substrate for protein chip detection. The XPS analysis of the surface elements of the two silicon-based wafers shows that the protein binds well to the pattern region of the wafer. The results of the protein test showed that, The sensitivity of protein detection on the surface of micropatterned SiO_2/Si silicon-based wafers and Au/SiO_2 silicon-based wafers is 10 渭 g / L. 3. The micropatterned SiO_2/Si wafers and micropatterned Au/SiO_2 wafers are used for DNA fluorescence detection. Both micropatterned silicon wafers have very strong fluorescence signals in the pattern area, but the background signals in the non-pattern region are very low, indicating that the SiO_2/Si chip and the Au/SiO_2 chip can completely eliminate the interference of the background signal when used for DNA detection. The SiO_2/Si chip and Au/SiO_2 chip can be used as the substrate for other DNA detection methods. The sensitivity of two kinds of wafers for DNA detection was investigated. The results showed that the detection limit of DNA on the surface of the micropatterned SiO_2/Si wafer was 10 pm, and the detection limit of DNA on the surface of the au-Si Sio _ 2 wafer was 100 pm 路cm ~ (- 1). 4. The thickness of protein film on micro-patterned SiO_2/Si silicon wafer was analyzed by ellipsometry and two-beam interferometric optical detection. The results of ellipsometry show that the average thickness of protein film on the surface of SiO_2/Si silicon wafer is 4.3 nm. The two-beam interferometric optical detection system was used to detect the signal output of rabbit anti-human IgG with different concentrations. The results showed that the thickness of the signal was increased by about 4.27 nm, which was consistent with the ellipsometry. This indicates that the micro-patterned SiO_2/Si silicon wafer can be used in the label-free optical detection system.
【学位授予单位】：湖南工业大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.0
【参考文献】
相关期刊论文 前8条
1 宋致堂;李国华;张霞;孔凡美;宋师霞;;椭圆偏振术在生物分子芯片研究中的应用[J];曲阜师范大学学报(自然科学版);2008年03期
2 孟艳丽,王战会,靳刚;A蛋白定向固定抗体用于椭偏光学生物传感器免疫检测[J];生物工程学报;2004年01期
3 费嘉,马文丽,郑文岭;蛋白质芯片在蛋白质组学研究中的作用[J];生命科学;2005年02期
4 王岸娜;刘小彦;吴立根;徐山宝;;蛋白质的提取、纯化及性质研究概述[J];中国食品工业;2008年06期
5 田文杰;路峻岭;刘玲玲;张福学;;切变振动型石英晶体谐振器的特点及其应用[J];压电与声光;2006年02期
6 石志松;朱惠忠;王文炎;;用于液相生物量检测的单片阵列QCM传感器[J];仪器仪表学报;2006年S2期
7 段宗涛;;石英晶体离子刻蚀微调原理与应用[J];中国高新技术企业;2010年21期
8 胡勇,曾雪梅,公衍道,隋森芳,赵南明;用椭圆偏振术研究血清白蛋白和纤维蛋白原在硅片及聚氨酯薄膜表面的吸附行为[J];中国生物医学工程学报;1998年01期
相关博士学位论文 前1条
1 周忠海;基于激光干涉原理的海洋灾害预警关键技术研究[D];中国海洋大学;2009年
相关硕士学位论文 前4条
1 陈晗;生物光盘传感器理论分析及初步实验研究[D];国防科学技术大学;2010年
2 张恒;基于光度法及混合法的自动椭偏仪研究[D];华南师范大学;2005年
3 刘斌;差分型SPR仪的搭建及用于万古霉素/抗体相互作用研究[D];中南大学;2007年
4 杨杰;两种双光束干涉型光纤传感器的研究[D];山东大学;2008年
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