基于DNA雜交的新型液晶傳感器及其應(yīng)用
發(fā)布時間：2018-10-08 14:06
【摘要】：近年來,DNA檢測廣泛應(yīng)用于臨床診斷、基因表達(dá)、農(nóng)業(yè)、食品安全以及環(huán)境監(jiān)測等領(lǐng)域,因此對DNA進(jìn)行快速、便捷、靈敏的檢測十分必要。傳統(tǒng)的DNA檢測方法有基于熒光的生物芯片技術(shù)、表面等離子共振(Surface Plasmon Resonance,SPR)等,但是這些檢測方法存在諸多不足,仍需改進(jìn)。近年來,具有獨(dú)特物理化學(xué)性質(zhì)的新型材料——液晶已經(jīng)引起研究人員的廣泛關(guān)注。液晶是介于液態(tài)與固態(tài)之間的一種物質(zhì)形態(tài),具有雙折射性、彈性應(yīng)變等特性,基底表面微小的變化即可引起液晶分子取向的變化,而液晶分子取向的變化可以轉(zhuǎn)變?yōu)楣鈱W(xué)信號,肉眼即可觀察,因此在DNA檢測中備受關(guān)注。本文基于DNA雜交和液晶的雙折射性,構(gòu)建了固態(tài)-液晶傳感界面,并在其基礎(chǔ)上對DNA雜交進(jìn)行了檢測;除此之外,本文還構(gòu)建了水-液晶界面,探究了不同表面活性劑對液晶分子排列的影響,并在其基礎(chǔ)上探索了單鏈DNA(ssDNA)對水-液晶界面的影響,為DNA雜交的檢測奠定了基礎(chǔ)。具體內(nèi)容如下:(1)利用ssDNA作為取向劑在固態(tài)-液晶界面構(gòu)建功能化基底。首先將三乙氧基硅基丁醛(TEA)修飾于固態(tài)基底表面,利用帶有醛基的TEA與修飾有氨基的ssDNA之間的反應(yīng),將ssDNA修飾在固態(tài)基底表面。探究了玻片在TEA中的修飾時間以及ss DNA在基底表面的表面覆蓋率對于功能化基底構(gòu)建的影響。(2)基于固態(tài)-液晶界面檢測DNA雜交的探索。采用上述方法構(gòu)建一種新型的利用ssDNA誘導(dǎo)液晶4-氰基-4’-戊基聯(lián)苯(5CB)垂直排列的功能化基底,通過優(yōu)化實(shí)驗(yàn)條件,ssDNA會誘導(dǎo)5CB分子垂直排列,偏光顯微鏡下觀察到均一的黑色,當(dāng)加入互補(bǔ)ssDNA之后,會發(fā)生雜交反應(yīng),擾亂5CB分子的排列,偏光顯微鏡下觀察到彩色紋理,從而達(dá)到檢測DNA的效果。在優(yōu)化實(shí)驗(yàn)條件(玻片在TEA溶液中的修飾時間為1h,ssDNA的最佳濃度為20 nM)下,該檢測方法的檢測限為0.1 nM,并且具有良好的特異性和重復(fù)性。(3)在水-液晶界面不同表面活性劑對5CB分子取向的影響。根據(jù)表面活性劑與5CB分子之間的相互作用,探討了在水-液晶界面不同表面活性劑(十八烷基三甲基溴化銨,十二烷基三甲基溴化銨,正辛基三甲基溴化銨,十二烷基硫酸鈉)對5CB分子取向的影響,探究了每種表面活性劑誘導(dǎo)5CB分子垂直排列的最佳濃度與響應(yīng)時間,鏈長不同的同類表面活性劑對5CB分子取向的影響,以及鏈長相同的不同類表面活性劑對5CB分子取向的影響。除此之外,我們還探究了在水-液晶界面ssDNA對5CB分子取向的影響,為后續(xù)DNA雜交的檢測提供了方法與條件。
[Abstract]:In recent years, DNA detection has been widely used in clinical diagnosis, gene expression, agriculture, food safety and environmental monitoring, so it is necessary to detect DNA quickly, conveniently and sensitively. Traditional DNA detection methods are based on fluorescence biochip technology, surface plasmon resonance (Surface Plasmon Resonance,SPR) and so on, but these detection methods have many shortcomings and still need to be improved. In recent years, liquid crystal, a new material with unique physical and chemical properties, has attracted wide attention. Liquid crystal is a kind of material form between liquid and solid. It has properties of birefringence, elastic strain and so on. The change of the orientation of liquid crystal molecules can be transformed into optical signals, which can be observed by the naked eye, so it has attracted much attention in DNA detection. Based on the birefringence of DNA hybridization and liquid crystal, the solid-liquid crystal sensing interface was constructed, and the DNA hybridization was detected on the basis of it. In addition, the water-liquid crystal interface was constructed in this paper. The effects of different surfactants on the arrangement of liquid crystal molecules were investigated, and the effects of single strand DNA (ssDNA) on the water-liquid crystal interface were explored, which laid a foundation for the detection of DNA hybridization. The main contents are as follows: (1) ssDNA was used as the orientation agent to construct the functional substrate at the solid-liquid crystal interface. Firstly, triethoxysilyl butyraldehyde (TEA) was modified on the solid substrate surface. SsDNA was modified on the solid substrate surface by the reaction between TEA with aldehyde group and ssDNA modified with amino group. The effects of glass modification time in TEA and ss DNA surface coverage on the construction of functional substrate were investigated. (2) Detection of DNA hybridization based on solid-liquid crystal interface. The above method was used to construct a novel functional substrate for the vertical alignment of liquid crystal 4-cyano-4keto-pentyl biphenyl (5CB) induced by ssDNA. The vertical arrangement of 5CB molecules was induced by optimizing the experimental conditions, and the uniform black was observed under polarizing microscope. When the complementary ssDNA is added, the hybridization reaction will occur, and the arrangement of 5CB molecules will be disturbed. The color texture can be observed under the polarizing microscope, thus the effect of DNA detection can be achieved. Under the optimized experimental conditions (the time of modification of glass slides in TEA solution is 1 h), the optimum concentration of DNA is 20 nM. The detection limit of this method is 0.1 nM, and has good specificity and repeatability. (3) the effect of different surfactants on the orientation of 5CB molecules at the water-liquid crystal interface. Based on the interaction between surfactants and 5CB molecules, different surfactants (octadecyl trimethylammonium bromide, dodecyl trimethyl ammonium bromide, octyl trimethylammonium bromide) at the water-liquid crystal interface were studied. The effect of sodium dodecyl sulfate on the orientation of 5CB molecules was investigated. The optimum concentration and response time of each surfactant to induce the vertical alignment of 5CB molecules, and the effects of the same surfactants with different chain length on the orientation of 5CB molecules were investigated. And the effects of different surfactants with the same chain length on the orientation of 5CB molecules. In addition, we also investigated the effect of ssDNA on the orientation of 5CB molecules at the water-liquid crystal interface, which provided the methods and conditions for the subsequent detection of DNA hybridization.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TP212;Q523
本文編號：2257041
[Abstract]:In recent years, DNA detection has been widely used in clinical diagnosis, gene expression, agriculture, food safety and environmental monitoring, so it is necessary to detect DNA quickly, conveniently and sensitively. Traditional DNA detection methods are based on fluorescence biochip technology, surface plasmon resonance (Surface Plasmon Resonance,SPR) and so on, but these detection methods have many shortcomings and still need to be improved. In recent years, liquid crystal, a new material with unique physical and chemical properties, has attracted wide attention. Liquid crystal is a kind of material form between liquid and solid. It has properties of birefringence, elastic strain and so on. The change of the orientation of liquid crystal molecules can be transformed into optical signals, which can be observed by the naked eye, so it has attracted much attention in DNA detection. Based on the birefringence of DNA hybridization and liquid crystal, the solid-liquid crystal sensing interface was constructed, and the DNA hybridization was detected on the basis of it. In addition, the water-liquid crystal interface was constructed in this paper. The effects of different surfactants on the arrangement of liquid crystal molecules were investigated, and the effects of single strand DNA (ssDNA) on the water-liquid crystal interface were explored, which laid a foundation for the detection of DNA hybridization. The main contents are as follows: (1) ssDNA was used as the orientation agent to construct the functional substrate at the solid-liquid crystal interface. Firstly, triethoxysilyl butyraldehyde (TEA) was modified on the solid substrate surface. SsDNA was modified on the solid substrate surface by the reaction between TEA with aldehyde group and ssDNA modified with amino group. The effects of glass modification time in TEA and ss DNA surface coverage on the construction of functional substrate were investigated. (2) Detection of DNA hybridization based on solid-liquid crystal interface. The above method was used to construct a novel functional substrate for the vertical alignment of liquid crystal 4-cyano-4keto-pentyl biphenyl (5CB) induced by ssDNA. The vertical arrangement of 5CB molecules was induced by optimizing the experimental conditions, and the uniform black was observed under polarizing microscope. When the complementary ssDNA is added, the hybridization reaction will occur, and the arrangement of 5CB molecules will be disturbed. The color texture can be observed under the polarizing microscope, thus the effect of DNA detection can be achieved. Under the optimized experimental conditions (the time of modification of glass slides in TEA solution is 1 h), the optimum concentration of DNA is 20 nM. The detection limit of this method is 0.1 nM, and has good specificity and repeatability. (3) the effect of different surfactants on the orientation of 5CB molecules at the water-liquid crystal interface. Based on the interaction between surfactants and 5CB molecules, different surfactants (octadecyl trimethylammonium bromide, dodecyl trimethyl ammonium bromide, octyl trimethylammonium bromide) at the water-liquid crystal interface were studied. The effect of sodium dodecyl sulfate on the orientation of 5CB molecules was investigated. The optimum concentration and response time of each surfactant to induce the vertical alignment of 5CB molecules, and the effects of the same surfactants with different chain length on the orientation of 5CB molecules were investigated. And the effects of different surfactants with the same chain length on the orientation of 5CB molecules. In addition, we also investigated the effect of ssDNA on the orientation of 5CB molecules at the water-liquid crystal interface, which provided the methods and conditions for the subsequent detection of DNA hybridization.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TP212;Q523
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