基于微圖案硅基晶片的生物檢測研究
發(fā)布時間：2019-03-13 08:46
【摘要】：生物芯片和微陣列是生物檢測技術(shù)的一個重要發(fā)展趨勢。在芯片制備中，選擇合適的基片材料與點陣排列，降低點陣與背景、點陣與點陣之間的信號干擾，是需要解決的課題之一。本文設(shè)計了微圖案SiO_2/Si硅基晶片和微圖案Au/SiO_2硅基晶片兩種基材，對其表面處理、生物檢測可行性以及在相關(guān)光學檢測方法中的應(yīng)用進行了探討，主要研究內(nèi)容如下： 1、微圖案SiO_2/Si硅基晶片的表面處理與修飾，采用濃H_2SO_4/H_2O_2、濃HCL/H_2O_2/H_2O、NH_4OH/H_2O_2/H_2O三種清洗液對其表面進行親水化處理，結(jié)果顯示H_2SO_4/H_2O_2溶液的處理效果最好，最佳處理時間為30分鐘。同時對微圖案SiO_2/Si晶片表面的共價修飾方法和疏水化處理方法進行了比較，結(jié)果表明，前者對蛋白質(zhì)的固定效果優(yōu)于后者。對微圖案Au/SiO_2晶片，采用MUA修飾其表面，接觸角測量結(jié)果表明，Au/SiO_2晶片表面已成功修飾MUA，可進一步與生物分子連接。 2．微圖案SiO_2/Si晶片和微圖案Au/SiO_2晶片用于蛋白質(zhì)熒光檢測。結(jié)果表明，兩種晶片固定人IgG底物的最佳濃度均為100mg/L，兩種晶片均在圖案區(qū)獲得相當強的熒光信號，而非圖案區(qū)背景信號很低，可作為基材用于蛋白質(zhì)芯片檢測。對兩種硅基晶片進行表面元素的XPS分析，證明蛋白質(zhì)已與晶片圖案區(qū)很好地結(jié)合。對蛋白質(zhì)的檢測結(jié)果表明，微圖案SiO_2/Si硅基晶片和Au/SiO_2硅基晶片表面的蛋白質(zhì)檢測靈敏度均為10μg/L。 3．微圖案SiO_2/Si晶片和微圖案Au/SiO_2晶片用于DNA熒光檢測。兩種微圖案硅基晶片均在圖案區(qū)獲得相當強的熒光信號，而非圖案區(qū)背景信號很低，表明SiO_2/Si晶片和Au/SiO_2晶片用于DNA檢測，可完全排除背景信號的干擾，確定了SiO_2/Si晶片和Au/SiO_2晶片可作為其它DNA檢測方法的芯片基材而進一步推廣應(yīng)用。分別考察了兩種晶片的DNA檢測靈敏度，結(jié)果表明，微圖案SiO_2/Si晶片表面的DNA檢測限為10pM，Au/SiO_2晶片表面的DNA檢測限為100pM。 4、采用橢偏檢測和雙光束干涉光學檢測法對微圖案SiO_2/Si硅基晶片表面蛋白質(zhì)膜層厚度進行分析。橢偏檢測的結(jié)果顯示，SiO_2/Si硅基晶片表面蛋白質(zhì)膜層的平均厚度為4.3nm；通過雙光束干涉光學檢測系統(tǒng)檢測不同濃度的兔抗人IgG所輸出的信號，，結(jié)果顯示其厚度增加約4.27nm，與橢偏檢測的結(jié)果相吻合。這表明微圖案SiO_2/Si硅基晶片可以應(yīng)用于免標記的光學檢測系統(tǒng)。
[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.
【學位授予單位】：湖南工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R318.0
本文編號：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.
【學位授予單位】：湖南工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R318.0
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