本文選題：摻雜量子點 + 室溫磷光�。� 參考：《山西師范大學(xué)》2017年碩士論文

【摘要】：生物分子的研究使人們從基因水平上了解某些疾病的發(fā)病機理,并且通過分子探針的設(shè)計對機體內(nèi)生物分子的含量進行痕量檢測,為各種疾病的發(fā)生提供一定的理論依據(jù),同時為臨床醫(yī)學(xué)中使用劑量的有效控制提供一定的理論指導(dǎo)。納米分子探針具備水溶性好、光化學(xué)穩(wěn)定、比表面積大等特性,提高了傳統(tǒng)生物分子探針的檢測性能。而室溫磷光(Room-temperature phosphorescence,RTP)量子點納米分子探針不但具備上述優(yōu)點,還可以避免生物體液背景熒光及散射光干擾,在進行生物樣品檢測時更精準(zhǔn)。本研究利用室溫磷光量子點的光學(xué)性質(zhì),將其以納米探針的形式對幾種重要的生物分子(DNA、細(xì)胞色素C、殼寡糖)進行檢測。主要研究內(nèi)容如下:(1)通過3-巰基丙酸(3-Mercaptopropionic Acid,MPA)包裹的Mn摻雜ZnS(Mn-ZnS)量子點(Quantum Dots,QDs)與亞甲基藍(lán)(Methylene Blue,MB)之間的靜電作用構(gòu)建了一種納米復(fù)合物,并將其應(yīng)用于DNA的痕量檢測。其檢測原理為:亞甲基藍(lán)通過靜電作用與Mn-ZnS QDs結(jié)合,并通過光誘導(dǎo)電子轉(zhuǎn)移(Photoinduced Electron Transfer,PIET)過程猝滅了Mn-ZnS QDs的RTP;隨著DNA的加入,亞甲基藍(lán)通過嵌插和靜電作用與DNA結(jié)合,并且亞甲基藍(lán)從Mn-ZnS QDs表面脫附,使得QDs的RTP恢復(fù);據(jù)此,建立了一種基于RTP性質(zhì)的DNA檢測方法。該方法的檢測范圍為0.2-20 mg/L,方法檢出限為0.113 mg/L。由于該DNA檢測方法是基于QDs的RTP,避免了來自生物體液的背景熒光及散射光的干擾,且無需復(fù)雜的樣品前處理過程,因此該方法適合于生物體液中DNA的痕量檢測。(2)利用水相合成法制備了MPA包裹的Mn-ZnS QDs,基于該QDs的RTP性質(zhì),構(gòu)建了一種靈敏、快速檢測細(xì)胞色素C(Cytochrome C,Cyt C)含量的新方法。該檢測方法利用QDs與Cyt C之間發(fā)生電子傳遞過程,使得QDs的RTP猝滅,從而對Cyt C的含量進行檢測。在pH=7.4的磷酸緩沖液中,Cyt C通過電子傳遞可以猝滅Mn-ZnS QDs在590 nm處的RTP,且在一定的范圍內(nèi)Cyt C濃度與RTP猝滅強度(P0/P)呈現(xiàn)良好的線性關(guān)系,線性范圍為0.1-10μM和10-40μM,相關(guān)系數(shù)為0.990和0.991,方法檢出限為0.062μM,該方法適用于Cyt C的痕量檢測。(3)通過MPA包裹的Mn-ZnS QDs與殼寡糖(chitosan oligosaccharide,COS)之間的靜電作用,構(gòu)建了一種精準(zhǔn)、高效檢測殼寡糖含量的新方法。該檢測方法利用QDs與殼寡糖之間的相互作用,使得QDs的RTP增強,從而對殼寡糖的含量進行檢測。在pH=7.4的磷酸緩沖液中,殼寡糖可以加強Mn-ZnS QDs在590nm處的RTP,且在一定的范圍內(nèi)殼寡糖濃度與RTP增強強度(P/P0)呈現(xiàn)良好的線性關(guān)系,線性范圍為0.06-10μg/mL,相關(guān)系數(shù)為0.988,方法檢出限為0.028μg/mL,該方法適用于殼寡糖的痕量檢測。總之,本文利用QDs的RTP性質(zhì),通過構(gòu)建納米探針對生物分子DNA、細(xì)胞色素C、殼寡糖的含量進行檢測,為分子生物學(xué)及各種疾病發(fā)生的研究提供了一定的理論依據(jù)。
[Abstract]:The study of biomolecules enables people to understand the pathogenesis of some diseases from the gene level, and to detect the content of biomolecules in the organism by the design of molecular probes, which provides a certain theoretical basis for the occurrence of various diseases. At the same time, it provides some theoretical guidance for effective control of dosage in clinical medicine. Nano-molecular probes have the characteristics of good water solubility, photochemical stability and large specific surface area, which improve the detection performance of traditional biomolecular probes. The room temperature phosphorescence (RTP) quantum dot nano-molecular probe not only has the advantages, but also avoids the interference of background fluorescence and scattering light, and is more accurate in the detection of biological samples. In this study, the optical properties of room temperature phosphorescent quantum dots (RQDs) were used to detect the DNA, cytochrome C and oligosaccharide of several important biomolecules in the form of nano-probe. The main contents of this study are as follows: (1) A nanocomposite was constructed by electrostatic interaction between Mn-doped ZnSn-ZnSn-ZnSn-ZnDs) and methylene blue methylene (MBB), which was encapsulated by 3-Mercaptopropionic Acid-MPA-encapsulated with 3-mercaptopropionic MPA. The nanocomposites were used for the determination of trace amounts of DNA. The detection principles are as follows: methylene blue binds to Mn-ZnS QDs by electrostatic interaction and quenches Mn-ZnS QDs through photoinduced Electron transfer via photoinduced electron transfer. With the addition of DNA, methylene blue binds to DNA through intercalation and electrostatic interaction. The methylene blue was desorbed from the surface of Mn-ZnS QDs to restore the RTP of QDs, and a DNA detection method based on RTP property was established. The detection range of the method is 0.2-20 mg / L, and the detection limit is 0.113 mg / L. Because the DNA detection method is based on QDs, it avoids the interference of background fluorescence and scattering light from biological body fluid, and does not need complicated sample pretreatment process. Therefore, this method is suitable for the trace detection of DNA in biological fluid. (2) based on the RTP properties of MPA, a new sensitive and rapid method for detecting the content of cytochrome C(Cytochrome Cy C was constructed by using the aqueous phase synthesis method to prepare Mn-ZnS QDsencapsulated with MPA. This method uses the electron transfer process between QDs and Cyt C to quench the RTP of QDs, so that the content of Cyt C is determined. In the phosphoric acid buffer of pH=7.4, Cyt C can quench the Mn-ZnS QDs at 590nm by electron transfer, and there is a good linear relationship between the concentration of Cyt C and the quenching intensity of RTP (P0 / P0) in a certain range. The linear ranges are 0.1-10 渭 M and 10-40 渭 M, the correlation coefficients are 0.990 and 0.991.The detection limit of the method is 0.062 渭 M. this method is suitable for the trace detection of Cyt C. This method is suitable for the detection of Cyt C by electrostatic interaction between Mn-ZnS QDs wrapped with MPA and chitosan oligosaccharides. A new method for the determination of chitosan oligosaccharide content. By using the interaction between QDs and oligosaccharides, the RTP of QDs was enhanced, and the content of chitosan oligosaccharide was detected. In the phosphoric acid buffer of pH=7.4, the chitosan oligosaccharide can enhance the Mn-ZnS QDs at 590nm, and the concentration of chitosan oligosaccharide has a good linear relationship with the enhancement strength of RTP in a certain range. The linear range is 0.06-10 渭 g / mL, the correlation coefficient is 0.988, and the detection limit is 0.028 渭 g / mL. This method is suitable for the trace determination of chitosan oligosaccharides. In conclusion, the content of DNA, cytochrome C and oligosaccharide in biomolecules were detected by using the RTP properties of QDs, which provided a theoretical basis for the study of molecular biology and the occurrence of various diseases.
【學(xué)位授予單位】：山西師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O471.1;R318

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