微梁陣列生化傳感器研制及檢測應用
發(fā)布時間：2018-08-30 10:45
【摘要】：微懸臂梁生化傳感技術是一項綜合微機電系統(tǒng)、生物化學以及力學等多項學科的新型分析方法。這種傳感技術以原子力顯微鏡技術為基礎,通過探測微懸臂梁偏轉等信號,對懸臂梁表面上發(fā)生的生化反應進行實時探測。該傳感技術具有靈敏度高、無須標記、實時原位等優(yōu)點,在生化分析等領域中具有廣泛的研究和應用前景。本文以微懸臂梁傳感技術為基礎,在已有的多種微懸臂梁傳感系統(tǒng)平臺的基礎上,搭建了能夠實時檢測包含八根微梁的陣列生化傳感系統(tǒng)。該系統(tǒng)利用光纖引導激光束作為微梁陣列探測光源,采用時序點亮照明微梁陣列的方式實現(xiàn)對微梁陣列的探測,有效地消除了懸臂梁信號中由溫度漂移、震動等因素帶來的噪聲,實現(xiàn)了高通量的檢測。隨后對系統(tǒng)進行了多項性能評估和優(yōu)化設計,有效地降低了信號噪聲、提高了檢測靈敏度,提升了系統(tǒng)的宜用性。最后在此傳感系統(tǒng)平臺上利用適配體成功的實現(xiàn)了對微囊藻毒素LR型的定量檢測。本論文的主要工作如下:搭建了以多束光纖引導激光束作為懸臂梁探測光源的微梁陣列傳感系統(tǒng),該系統(tǒng)能夠實時檢測包含八根懸臂梁的微梁陣列的偏轉信息。對系統(tǒng)的整體結構、器件的使用進行了詳細介紹;對傳感系統(tǒng)的光路進行優(yōu)化選擇,有效地減弱了雜散反射光的影響,使激光對準更加方便;設計了結構、體積合理的反應池,節(jié)約了樣品、方便了實驗操作。設計制作了能夠與傳感系統(tǒng)搭配使用的微梁陣列芯片。對研制的微梁陣列傳感系統(tǒng)的信號噪聲進行了分析,對激光掃描周期對信號噪聲的影響進行研究,確定每個掃描周期內單個激光工作時長不少于400ms;使用商品化的微梁陣列,利用雙材料梁的溫升效應,對傳感系統(tǒng)響應一致性進行評估,測得的各懸臂梁溫度響應靈敏度相對偏差不大于7.8%, 一致性良好。使用免疫抗原抗體檢測方法對銅離子和瘦肉精進行定量檢測,一致性良好,并能夠體現(xiàn)微梁陣列中參考組與實驗組的對比意義。對水溶液中的汞離子進行定量檢測,在同一濃度下檢測結果一致性良好,實驗中檢測濃度最低為0.1ng/mL。對制作的微梁陣列芯片測試和使用。測得懸臂梁曲率半徑約為15mm,能夠保證探測實驗對使用懸臂梁平直度的要求;制作的微梁陣列在溫度響應一致性測試中,各懸臂梁偏轉量與溫度呈良好的線性關系,擬合系數(shù)均大于0.99,而且各懸臂梁的溫度響應靈敏度相對偏差不大于5.0%,表現(xiàn)出良好的性能。使用制作的微梁陣列對汞離子進行定量檢測并與商品化陣列梁的檢測結果進行對比,1ng/mL的汞離子使制作的微梁產(chǎn)生偏轉量為21.0nm,0.2ng/mL的汞離子是商品化的微梁產(chǎn)生偏轉量為39.0nm,制作的微梁陣列剛度偏大,檢測靈敏度較低。利用單梁傳感平臺和修飾了特異性適配體的微梁對有機磷農(nóng)藥甲拌磷進行檢測。利用微梁陣列傳感系統(tǒng)和修飾了特異性適配體的微梁陣列對微囊藻毒素LR型進行定量檢測,檢測濃度范圍為1-500ng/mL,理論檢測極限為0.6ng/mL,提出的該基于適配體的微梁傳感方法能夠識別自來水中的微囊藻毒素LR型,并能夠區(qū)別微囊藻毒素的異構體專門用于微囊藻毒素LR型的檢測。
[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.
【學位授予單位】：中國科學技術大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：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.
【學位授予單位】：中國科學技術大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：TP212
【參考文獻】
相關期刊論文 前10條
1 鄔林;周夏榮;吳尚犬;王萍;張青川;伍小平;;新型微梁陣列生化傳感器的研制[J];分析化學;2012年04期
2 魏松紅;逄若霖;王，
本文編號：2212835
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