本文關(guān)鍵詞： 氧化鋅量子點 炭疽芽孢桿菌 吡啶二羧酸鈣 雙比色熒光傳感器　出處：《鄭州大學》2017年碩士論文　論文類型：學位論文

【摘要】：氧化鋅量子點是一種新型的熒光納米傳感器,其顆粒粒徑小、發(fā)光強度高、光穩(wěn)定性好、生物毒性低,制備簡便及生產(chǎn)成本低等優(yōu)異特性,同時具有水溶性在生物熒光成像,生物熒光探針及載藥等許多領(lǐng)域具有廣闊的應用前景。炭疽芽孢桿菌作為潛在的生化武器嚴重威脅人類的生命健康,目前探測其存在的方法多種多樣,而鑭基熒光光譜法被廣泛推廣,該方法基于芽孢桿菌的生物標志物Ca DPA可有效敏化稀土離子的熒光發(fā)射,同時稀土離子具有窄的發(fā)射譜、發(fā)光壽命長的原理。銪(Eu)和鋱(Tb)是熒光傳感器中常用的鑭系元素離子,它們具有強的熒光,長的熒光壽命及尖銳的線狀發(fā)射帶。Eu3+的最強發(fā)射位于616 nm,該本征發(fā)射將不會受到530 nm處的Zn O QDs熒光發(fā)射的干擾,而544 nm處的Tb3+的本征發(fā)射峰與Zn O QDs的發(fā)射帶有重疊,同時Eu3+的紅色熒光也易于肉眼觀察。再次銪基熒光傳感器可有效降低芳香族化合物與Tb3+的結(jié)合而造成的假陽性結(jié)果。迄今為止,以氧化鋅量子點作為內(nèi)在參比熒光檢測炭疽芽孢桿菌研究尚未報道。綜上我們將選用銪離子與氧化鋅量子點結(jié)合,銪離子作為傳感單元,氧化鋅量子點作為參比單元,以制備性能優(yōu)異的雙比色熒光傳感器,實現(xiàn)對炭疽芽孢桿菌的快速,靈敏高選擇性的檢測。本文所取得的研究成果如下:1)成功制備了六方纖鋅礦結(jié)構(gòu)的氧化鋅量子點,其平均顆粒粒徑~5 nm,具有良好的水溶性,強的黃光發(fā)射,高量子產(chǎn)率(31.98%)及良好的發(fā)光穩(wěn)定性。同時氧化鋅量子點本身具有較低的生物毒性,制備簡便,生產(chǎn)成本低等優(yōu)質(zhì)的特性。該氧化鋅量子點表面帶有-NH2基團易于表面改性以設計開發(fā)性能優(yōu)異的納米熒光探針,在生物醫(yī)學成像、疾病診斷和治療等方面起著至關(guān)重要的作用。2)成功制備一種銪修飾的氧化鋅量子點納米結(jié)構(gòu)(Zn O/Eu),用于快速、靈敏的雙比色檢測炭疽芽孢桿菌的存在。具有高強黃色熒光發(fā)射的氧化鋅量子點作為一種內(nèi)在參比,銪離子(Eu3+)螯合在氧化鋅量子點的表面作為信號報告單元。當銪離子與芽孢桿菌的生物標志物Ca DPA配位后其紅色熒光強度將顯著增強,而氧化鋅量子點的黃色熒光卻保持不變,因此增加的Ca DPA濃度可以導致Zn O/Eu雜化納米結(jié)構(gòu)的兩種熒光強度比的變化。同時由于制備的氧化鋅量子點的吸收帶延伸到深紫外區(qū),覆蓋DPA在280 nm處的吸收峰,可實現(xiàn)氧化鋅量子點與DPA在同一激發(fā)波長(280 nm)下的共同激發(fā),所以該ZnO/Eu納米結(jié)構(gòu)有望實現(xiàn)對CaDPA的雙比色熒光檢測。3)該傳感器時間響應曲線展示傳感反應可在~8 s內(nèi)完成,從而能夠快速檢測炭疽芽孢桿菌孢子的存在。在同一激發(fā)波長下,該傳感器中氧化鋅量子點和Eu3+的發(fā)光強度比值(I616/I530)與Ca DPA在0~4μM濃度范圍內(nèi)呈良好的線性關(guān)系,使其能實現(xiàn)對微量濃度的Ca DPA進行定量測定,同時通過IUPAC方法計算Zn O/Eu納米結(jié)構(gòu)對Ca DPA的檢測極限為3 n M,此檢測極限遠低于人體對炭疽芽孢桿菌感染劑量(60μM)。比率熒光測試也有效降低外界環(huán)境的干擾,以提升檢測結(jié)果的準確性。增大Ca DPA的濃度該傳感器在紫外燈照射下溶液顏色由黃色過渡到紅色,因人眼對顏色變化敏感,該傳感器也利于裸眼檢測Ca DPA的濃度。最后該比率熒光傳感器對Ca DPA顯示出比其它芳族配體,氨基酸和常見細胞離子具有顯著的選擇性。因此快速地響應、良好的靈敏度及選擇性使得Zn O/Eu納米結(jié)構(gòu)在檢測炭疽細菌中提供了有效便捷的方式,在臨床分析中具有巨大的應用潛力。
[Abstract]:Zinc Oxide is a new type of quantum dot fluorescent nano sensor, its small particle size, high luminous intensity, good light stability, low toxicity and low production cost, simple preparation and excellent properties, it is soluble in water in biological fluorescence imaging, and has broad application prospects in biological fluorescent probes and drug loading and many other fields Bacillus anthracis. As a potential biological weapon serious threat to human life and health, the current detection methods are varied, and the lanthanum based fluorescence spectroscopy has been widely popularized, the method based on Bacillus biomarker fluorescence emission Ca DPA can effectively sensitized, and rare earth ions having narrow emission spectra. The principle of luminescence and long life. (Eu) europium and terbium (Tb) fluorescent sensor lanthanide ions commonly used, they have strong fluorescence, long fluorescence lifetime and sharp linear emission band.Eu3+ The strongest emission at 616 nm, the intrinsic emission will be at 530 nm Zn O QDs fluorescence emission interference, and the intrinsic emission peak and Zn O QDs emission at 544 nm Tb3+ with red fluorescence overlap, while the Eu3+ is easily observed. Again based europium fluorescent sensors can have false positive the effect of reduced binding of aromatic compounds and Tb3+ caused. So far, Zinc Oxide quantum dots as internal reference fluorescence detection of Bacillus anthracis research have not been reported yet. So we will choose europium ion and Zinc Oxide quantum dots with europium ion as a sensing unit, Zinc Oxide quantum dots as a reference cell, producing excellent performance dual color fluorescence sensor for realization of Bacillus anthracis, rapid, sensitive and highly selective detection. The results of this study are as follows: 1) Zinc Oxide six quantum wurtzite structure were prepared Point, the average particle size of ~5 nm, has good water solubility, strong yellow emission, high quantum yield (31.98%) and good luminescent stability. At the same time Zinc Oxide quantum dot itself has low toxicity, easy preparation, low production cost and good quality. The surface of the quantum dots with Zinc Oxide -NH2 group of easy surface modification for the design of nano fluorescent probe excellentperformance, in biomedical imaging, plays a crucial role in.2 disease diagnosis and treatment) were prepared by Zinc Oxide quantum dot nanostructures for europium were successfully modified (Zn, O/Eu) for rapid, sensitive double colorimetric detection of Bacillus anthracis there is Zinc Oxide quantum dots. High strength yellow fluorescence emission as an internal reference, europium (Eu3+) chelate on the surface of Zinc Oxide quantum dots as a signal unit. When the report of europium ion and Bacillus biomarkers The red fluorescence intensity will be significantly enhanced Ca DPA markers after coordination, while Zinc Oxide yellow fluorescent quantum dots has remained unchanged, so Ca increased DPA concentration can lead to changes of two kinds of fluorescence intensity of Zn O/Eu hybrid nanostructures. At the same time as Zinc Oxide than quantum dots prepared by extending to the deep ultraviolet absorption band DPA, covered in the peak of 280 nm, Zinc Oxide and DPA can realize quantum dots at the same excitation wavelength (280 nm) under the common excitation, so the ZnO/Eu nano structure is expected to achieve the CaDPA dual color fluorescence detection.3) the sensor response time curve display sensing reaction can be completed in ~8 s thus, rapid detection of Bacillus anthracis spores. At the same excitation wavelength, emission intensity ratio of quantum dots and the Zinc Oxide Eu3+ sensor (I616/I530) and Ca DPA in 0~4 M concentration range showed a good linear relationship The system can realize the trace concentrations of Ca DPA were measured and calculated by the IUPAC method, the detection limit of Zn O/Eu nano structure of Ca DPA 3 N M, the detection limit is far lower than the human dose infection of Bacillus anthracis (60 M). The fluorescence test also effectively reduce interference ratio of low external environment in order to improve the accuracy of the test results. The increase of the concentration of Ca DPA in the sensor solution under the irradiation of ultraviolet light color from yellow to red transition, because of the human eye to color sensitive to changes in concentration of the sensor to the naked eye detection of Ca DPA. Finally, the ratio of fluorescence sensor for Ca DPA than other aromatic ligand, amino acids and common cell ions have significant selectivity. Therefore, fast response, good sensitivity and selectivity makes Zn O/Eu nanostructures in the detection of anthrax bacteria provides effective and convenient way in clinical analysis It has great potential for application.

【學位授予單位】：鄭州大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O657.3

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