發(fā)布時(shí)間：2018-04-05 07:37

  本文選題：碳化硅量子點(diǎn)　切入點(diǎn)：化學(xué)腐蝕法　出處：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：活體細(xì)胞長時(shí)程熒光成像示蹤及其細(xì)胞精細(xì)結(jié)構(gòu)顯示為生命科學(xué)研究提供了重要的技術(shù)支撐,可以用于細(xì)胞生長規(guī)律、轉(zhuǎn)基因?qū)ほ櫦皠?dòng)物疾病防治等領(lǐng)域,但傳統(tǒng)的熒光材料(如熒光蛋白、熒光染料、CdX量子點(diǎn)等)均不適合于活體細(xì)胞的長時(shí)程熒光成像與示蹤。近年來新型碳化硅量子點(diǎn)(SiC-QDs)由于具有優(yōu)良的生物相容性及光學(xué)性能,可以作為一種活體細(xì)胞長時(shí)程熒光成像示蹤材料,被材料界與生物學(xué)界學(xué)者所矚目。但到目前為止,化學(xué)腐蝕法可控制備SiC-QDs工藝與性能優(yōu)化、成型機(jī)制、活體細(xì)胞標(biāo)記機(jī)制等問題還沒有得到系統(tǒng)研究。本文針對(duì)這些問題進(jìn)行了一系列研究,結(jié)論如下:(1)以自蔓延燃燒合成的均質(zhì)納米SiC顆粒為原料,通過化學(xué)多重腐蝕,機(jī)械研磨結(jié)合超聲破碎,超重力場(chǎng)層析剪裁等工藝參數(shù)優(yōu)化,系統(tǒng)研究了SiC量子點(diǎn)成型過程中微觀形貌的演變規(guī)律及制備工藝參數(shù)對(duì)SiC量子點(diǎn)光學(xué)特性的影響規(guī)律,揭示了量子點(diǎn)制備工藝-微觀結(jié)構(gòu)-光學(xué)特性間的相互關(guān)聯(lián)機(jī)制,形成了一種SiC量子點(diǎn)標(biāo)記材料可控制備工藝,其步驟為:納米均質(zhì)SiC自蔓延燃燒合成→配制混合腐蝕液→進(jìn)行SiC顆粒腐蝕→降酸→烘干→機(jī)械研磨→SiC顆粒二次腐蝕→降酸→烘干→機(jī)械研磨→超聲空化破碎→高速離心層析剪裁→SiC量子點(diǎn)收集。(2)研究了新型工藝可控制備SiC量子點(diǎn)的熒光特性及其致病鐮刀菌活體細(xì)胞標(biāo)記強(qiáng)度與機(jī)制。結(jié)果表明:當(dāng)激發(fā)光為340 nm時(shí),SiC量子點(diǎn)光致發(fā)光強(qiáng)度最大,隨激發(fā)光波長增加,發(fā)射波長發(fā)生紅移,具有較高的斯托克斯位移。由于熒光發(fā)射可以全色調(diào)諧,可實(shí)現(xiàn)近紫外或近紅外熒光檢測(cè),實(shí)現(xiàn)了自發(fā)熒光細(xì)胞的有效檢測(cè)與定量分析。對(duì)致病鐮刀菌活體細(xì)胞SiC量子點(diǎn)熒光標(biāo)記機(jī)制的研究結(jié)果表明,量子點(diǎn)通過網(wǎng)格蛋白依賴的內(nèi)吞方式進(jìn)入活體細(xì)胞內(nèi)部,并均勻分布,從而實(shí)現(xiàn)了穩(wěn)定熒光標(biāo)記。另外基于實(shí)驗(yàn)結(jié)果與理論分析,提出了致病鐮刀菌活體細(xì)胞SiC量子點(diǎn)熒光標(biāo)記模型。(3)研究了量子點(diǎn)熒光標(biāo)記對(duì)致病鐮刀菌自身生理生長的影響,得出標(biāo)記后菌株在生長速度、產(chǎn)孢與產(chǎn)色素能力均未表現(xiàn)出抑制現(xiàn)象,與對(duì)照組幾乎同步生長,證明了碳化硅量子點(diǎn)具有優(yōu)良的生物相容性與熒光強(qiáng)度穩(wěn)定性;最后借助量子點(diǎn)的熒光標(biāo)記動(dòng)態(tài)示蹤技術(shù),對(duì)尖孢鐮刀菌致病機(jī)制進(jìn)行了研究,并提出了致病性尖孢鐮刀菌引起的植物枯萎病致病機(jī)理。
[Abstract]:In vivo, long term fluorescence imaging and fine cell structure display provide important technical support for life science research, and can be used in cell growth law, transgenic tracing, animal disease prevention and treatment, and so on.However, traditional fluorescent materials (such as fluorescent proteins, fluorescent dye CdX quantum dots, etc.) are not suitable for long term fluorescence imaging and tracer of living cells.In recent years, new silicon carbide quantum dots (SiC-QDss) have been widely used as a long time fluorescence imaging tracer material for their excellent biocompatibility and optical properties.However, so far, the process and performance optimization, molding mechanism, in vivo cell labeling mechanism of controlled preparation of SiC-QDs by chemical corrosion method have not been systematically studied.In this paper, a series of studies have been carried out to solve these problems. The conclusions are as follows: (1) using homogenous SiC particles synthesized by self-propagating combustion as raw materials, mechanical grinding combined with ultrasonic crushing is carried out by chemical multi-corrosion.Optimization of processing parameters such as super gravity field chromatography trimming and so on. The evolution law of microstructure and the influence of preparation parameters on the optical properties of SiC quantum dots were studied systematically in the process of forming SiC quantum dots.The correlation mechanism between the preparation process, microstructure and optical properties of quantum dots was revealed, and a controllable preparation process of SiC quantum dots was formed.The steps are as follows: Nano-homogenous SiC self-propagating combustion synthesizer is used to prepare mixed corrosion solution for SiC particle corrosion reduction and acid-reducing drying and mechanical grinding of SiC particles secondary corrosion / drying machine grinding and ultrasonic cavitation crushingThe fluorescence characteristics of SiC quantum dots prepared by a new technique and the labeling intensity and mechanism of SiC quantum dots in vivo were studied.The results show that when the excitation light is 340 nm, the photoluminescence intensity of sic QDs is the highest. With the increase of excitation wavelength, the emission wavelength is red-shifted and has a high Stokes shift.Because the fluorescence emission can be tuned in full color, the near ultraviolet or near infrared fluorescence detection can be realized, and the effective detection and quantitative analysis of autofluorescence cells can be realized.The fluorescence labeling mechanism of SiC quantum dots in vivistic cells of Fusarium sp. was studied. The results showed that quantum dots entered the living cells through the endocytosis mode dependent on grid proteins and distributed evenly, thus realizing stable fluorescence labeling.In addition, based on the experimental results and theoretical analysis, a fluorescence labeling model of SiC quantum dots in vivistic cells of Fusarium disease was proposed. The effects of quantum dot fluorescence labeling on the physiological growth of pathogenic Fusarium were studied, and the growth rate of the labeled strain was obtained.The ability of spore production and pigment production showed no inhibition, and almost synchronized growth with the control group, which proved that silicon carbide quantum dots had excellent biocompatibility and fluorescence stability.The pathogenetic mechanism of Fusarium oxysporum was studied and the pathogenetic mechanism of Fusarium oxysporum was put forward.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O613.72;TB383.1

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