【作者】 王亮；

【导师】 毕红；

【作者基本信息】 安徽大学 ， 有机化学， 2010， 硕士

【摘要】 近年来,生物成像不仅仅作为一种研究工具而且在预防和监测各种疾病中也起着重要作用。正是如此,生物成像引起了国内外科研工作者的广泛关注,尤其是细胞成像,它需要荧光染料足够小能进入细胞而且不影响细胞的正常代谢活动。但常用的有机荧光染料通常是疏水甚至是不溶于水的,这极大地限制了其在生物领域的应用,只有少数种类的有机分子可用于生物标记,如罗丹明及其衍生物等；而其它一些有机染料,如花,虽然具有高的荧光量子产率和荧光稳定性,但目前只限于用在有机光电器件中。本论文的目的是将这些疏水的有机荧光染料制备成尺寸适宜的纳米粒子,将这些有机荧光纳米粒子与细胞一起共培养,使这些纳米粒子被细胞内吞并停留在特定的细胞器内,研究它们的细胞毒性以及是否具备细胞成像的功能。本文制备了有机荧光染料N,N-双(2,6-二异丙基苯胺)-1,6,7,12-四氯花-3,4,9,10-四羧酸二酰亚胺(PDI),然后在自制的氧化铝模板中制备了其纳米管,初步研究了PDI纳米管的荧光性能及其在生物领域的应用；另外,采用原位聚合的方法制备了PDI/Ag/PMMA复合纳米材料,并研究了复合材料的荧光性能,具体内容如下：1.利用负压沉积法在氧化铝(AAO)模板中制备了PDI纳米管,其中AAO模板是通过电化学阳极氧化法制备的,其外径及长度可控。通过扫描式电子显微镜(SEM)、高分辨透射电子显微镜(HR-TEM)和Zeta电位仪对PDI纳米管的形貌、表面电性等进行了表征,结果表明制备的PDI纳米管长度分布在几百纳米到几微米之间,平均直径在60 nm左右,非晶,表面带负电；紫外可见近红外分光光度计(UV-vis)和荧光光谱仪(PL)等测试结果证明纳米管的分子聚集态为J-聚集体。选取HepG-2细胞为模型,使其与PDI纳米管共培养,经过2 h后发现PDI纳米管就能被HepG-2细胞摄入,同时显现出很好的成像效果。利用四唑盐比色法(MTT)证实PDI纳米管具有很好的生物相容性,HepG-2细胞与合适浓度(小于60μg/mL)的PDI纳米管共培养72 h后其存活率仍高达90%。2.以聚乙烯比咯烷酮(PVP)为软模板,通过多元醇还原法制备了银纳米粒子(银纳米线和银纳米颗粒),SEM、HR-TEM、Ⅹ射线衍射(XRD)等测试结果表明所制备的银纳米粒子属于面心立方结构,银纳米线长径比在200左右,银纳米粒子粒径在50-500 nm之间。通过原位聚合法合成了PDI/Ag/PMMA复合纳米材料并通过TGA和HR-TEM研究了复合材料的结构性能；荧光显微镜研究了复合材料的荧光性能。该研究为制备新型复合荧光材料提供了一条可行途径。更多还原

【Abstract】 Bio-imaging especially live cell imaging has received considerable attention not only as a research tool, but also in the prevention and early diagnosis of various diseases. Live cell imaging requires the size of the fluorescent dyes is small enough to enter the cells and then be visualized easily without affecting the normal metabolism behavior of the cells. However, organic fluorescent dyes are generally hydrophobic or even insoluble, which greatly limits their application in biological systems, and so far only a few kinds of organic fluorescent dyes have been used for bio-imaging, such as rhodamine and its derivatives. Because of such concern of hydrophobic problems, other attractive dyes, such as perylene with high quantum yield has been widely used in organic electroluminescence devices but not for bio-imaging. The intention of this paper is to prepare nanoparticles of such hydrophobic organic dyes in appropriate size, as the fluorescent dye nanoparticles are incubated with the cells for a sufficient time, they can be uptaken by the cells and then locate in the certain organelles, the cytotoxicity and live cell imaging function of the organic fluorescent nanoparticles will be studied for the purpose of providing a new effective bio-imaging method by using organic fluorescent nanoparticles.In this paper, N, N’-bis-(2,6-diisopropylphenyl)-1,6,7,12-tetrachloroperylene-3,4,9,10-tetracarboxylic acid diimide (PDI) was synthesized, and the PDI nanotubes were prepared within the nanopores of anodic alumina templates (AAO). The fluorescent properties of PDI nanotubes were investigated and its application in live cell imaging was investigated primarily. In addition, PDI/Ag/PMMA nanoncomposite was successfully prepared by using an in-situ polymerization method and its fluorescent property was also studied. The detailed works can be summarized as follows:1. PDI nanotubes were successfully prepared within the nanopores of AAO templates. The AAO template, with which the pore size and length can be tuned easily, was prepared in lab by aluminum electrochemical oxidizing. The morphology, structure and surface potential of the PDI nanotubes were characterized by using SEM, HRTEM and Zeta potential meter, respectively. The results indicated that the nanotubes were in an amorphous phase with negative surface potential, and they had an average outer diameter of about 60 nm, with the lengths ranging from several hundred nanometers to several micrometers. The optical properties of the nanotubes were measured by using UV-visible near-infrared spectrophotometer (UV-vis) and fluorescence spectrometer (PL), which indicated that the nanotubes were formed by head-to-tail J-aggregates. It is noteworthy the PDI nanotubes showed high fluorescent property. In order to observe its bio-imaging effect, HepG-2 cells were chosen to be incubated with the PDI nanotubes, it was found that large numbers of the nanotubes entered into the cells after 2 hours co-incubation and then the cells could be visualized obviously by fluorescent microscopy observation. Moreover, the cytotoxicity of the PDI nanotubes was evaluated by MTT assay and the results have demonstrated that the cell viability of the HepG-2 cells exposed to effective concentration (≤60μg/mL) of nanotubes for 72 h was still as high as 90%.2. Silver nanowires and nanoparticles had been prepared by polyol reduction in the presence of poly (vinylpyrrolidone). The morphology and structure of Ag nanowires and nanoparticles were characterized by using SEM, XRD and HRTEM. The results showed that both of them were highly crystalline, but the Ag nanowires had a maximum aspect ratio of 200, while the Ag nanoparticles had a size distribution ranging from 50 to 500 nm. PDI/Ag/PMMA nanocomposites were synthesized by means of in-situ polymerization. The structure and fluorescent property of the nanocomposites were investigated by using TGA, HR-TEM and the fluorescent microscopy. This initial work did not result in a direct surface modification of Ag nanoparticles by organic PDI molecules, but the research author has done deserves further studies.更多还原