本文選題：聚集誘導(dǎo)發(fā)光　切入點(diǎn)：殼聚糖　出處：《浙江大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：熒光探針或化學(xué)分子熒光傳感器是一類能夠選擇性結(jié)合特定物質(zhì)的分子體系,通過紫外吸收、熒光發(fā)射光譜或氧化還原電位等變化來實(shí)現(xiàn)對(duì)靶向分子定性或定量檢測,具有操作簡便、靈敏度高等優(yōu)點(diǎn),被廣泛應(yīng)用于生命科學(xué)、環(huán)境科學(xué)等領(lǐng)域。然而,傳統(tǒng)的熒光分子往往存在不可忽視的缺點(diǎn),如隨濃度變化的stokes位移導(dǎo)致熒光強(qiáng)度的非線性變化、高濃度或聚集時(shí)發(fā)生熒光淬滅現(xiàn)象等,大大限制了其應(yīng)用范圍。而具有聚集誘導(dǎo)發(fā)光(AIE)特性的熒光分子針對(duì)性地解決了這一難題,并因其敏感度高、光譜隨濃度變化無漂移等優(yōu)異特性使之在熒光檢測各領(lǐng)域得到極大應(yīng)用。在具有AIE特性熒光探針被廣泛探究的同時(shí),水溶性AIE探針的需求日益迫切。殼聚糖富含氨基、羧基,是自然界僅次于纖維素的第二大天然可再生高分子材料,具有生物可降解性、生物相容性、易與金屬離子絡(luò)合等優(yōu)良性能,在生物醫(yī)藥、環(huán)境檢測等領(lǐng)域具有廣泛應(yīng)用。本論文將四苯基乙烯(TPE)熒光分子標(biāo)記在殼聚糖及其衍生物分子鏈上,制得具有AIE特性的殼聚糖基熒光分子,繼而拓展其在生物、環(huán)境監(jiān)測領(lǐng)域中的應(yīng)用。主要研究成果如下:1)采用均相反應(yīng)法在乙酸/甲醇溶劑體系中對(duì)殼聚糖進(jìn)行琥珀酰化改性,繼而標(biāo)記熒光分子,成功制備TPE-NSCS,并以之為檢測探針應(yīng)用于細(xì)胞長周期示蹤。1HNMR、熒光光譜(FL)、表面Zeta電位、TEM等測試結(jié)果表明成功制備表面負(fù)電性的水溶性佳、穩(wěn)定性好的AIE探針TPE-NSCS;用已制得的TPE-NSCS對(duì)宮頸癌細(xì)胞(HeLa)進(jìn)行染色,MTT結(jié)果證明其無細(xì)胞毒性,熒光顯微鏡照片及共聚焦顯微鏡(CLSM)圖片表明隨著染色時(shí)間增加,探針成功進(jìn)入細(xì)胞內(nèi)部,成像效果良好,且可長周期示蹤至30代。2)利用2,3-環(huán)氧丙基三甲基氯化銨對(duì)殼聚糖進(jìn)行季銨化改性,繼而標(biāo)記熒光分子,成功制備TPE-TMC,并以之為檢測探針應(yīng)用于細(xì)胞長周期示蹤。1HNMR、FL光譜、表面Zeta電位、TEM等測試結(jié)果表明成功制備表面正電性的水溶性佳、穩(wěn)定性好的AIE探針TPE-TMC;用已制得的TPE-TMC對(duì)HeLa進(jìn)行染色,MTT結(jié)果證明其無細(xì)胞毒性,熒光顯微鏡照片及CLSM圖片表明隨著染色時(shí)間增加,探針快速進(jìn)入細(xì)胞內(nèi)部,成像效果良好,且可長周期示蹤至25代。該優(yōu)異性能使之在藥物代謝檢測、細(xì)胞示蹤等領(lǐng)域具有良好而廣泛的應(yīng)用前景。3)成功制得具有AIE特性的TPE-CS,并以之為熒光探針研究了其與Cu(Ⅱ)之間相互作用。采用FL光譜、紫外吸收光譜、TEM等分析方法對(duì)其作用過程進(jìn)行表征,FL光譜表明,TPE-CS與不同金屬離子相互作用其熒光強(qiáng)度變化不同,且唯有與Cu(Ⅱ)作用呈現(xiàn)熒光光強(qiáng)減弱趨勢,且降低的熒光強(qiáng)度與Cu(Ⅱ)濃度在一定范圍內(nèi)呈現(xiàn)良好的線性關(guān)系,可用于定量檢測Cu(Ⅱ)。
[Abstract]:Fluorescence probe or chemical molecular fluorescence sensor is a kind of molecular system which can selectively bind to a specific substance. It can detect the target molecule qualitatively or quantitatively by the changes of ultraviolet absorption, fluorescence emission spectrum or redox potential. With the advantages of simple operation and high sensitivity, it has been widely used in life science, environmental science and other fields. However, traditional fluorescent molecules often have disadvantages that can not be ignored. For example, the shift of stokes with concentration leads to the nonlinear change of fluorescence intensity, and the phenomenon of fluorescence quenching occurs at high concentration or aggregation, etc. The fluorescence molecules with the characteristic of agglomeration induced luminescence (AIEs) have solved this problem pertinently, and because of its high sensitivity, The spectral properties such as no drift with concentration make it widely used in various fields of fluorescence detection. While the fluorescence probe with AIE characteristics has been widely explored, the demand of water-soluble AIE probe is increasingly urgent. Chitosan is rich in amino, carboxylic groups. Nature is the second largest natural renewable polymer material after cellulose, with biodegradability, biocompatibility, easy to complex with metal ions and other excellent properties in biomedicine, In this paper, the fluorescent molecular markers of tetraphenylethylene tetrachloride (TPE) were labeled on the molecular chain of chitosan and its derivatives, and chitosan based fluorescent molecules with AIE properties were prepared, and then their biological properties were expanded. Applications in the field of environmental monitoring. The main research results are as follows: (1) the succinylation of chitosan in acetic acid / methanol solvent system was carried out by homogeneous reaction method, and then fluorescent molecules were labeled. TPE-NSCSs were successfully prepared and used as detection probes for cell long cycle tracer. 1HNMRs, fluorescence spectra and Zeta potential Tem. The results showed that the water solubility of surface negative electricity was good. The stable AIE probe TPE-NSCS, which was stained with TPE-NSCS, showed no cytotoxicity. Fluorescence microscopy and confocal microscopy images showed that the probe successfully entered the cell interior with the increase of staining time. The imaging effect is good, and can be traced to 30 generations. 2) Quaternary ammonium chloride was used to modify chitosan and then label fluorescent molecules. TPE-TMC was successfully prepared and used as a probe for cell long period tracer. 1HNMRFL spectra. The results of Zeta potential Tem on the surface showed that the water solubility of the positive surface was good. The stable AIE probe TPE-TMC.The HeLa was stained with TPE-TMC and proved to be non-cytotoxic. Fluorescence microscope and CLSM images showed that the probe quickly entered the cell interior with the increase of dyeing time, and the imaging effect was good. And can be traced for a long period to 25 generations. TPE-CSs with AIE properties were successfully prepared and their interaction with Cu2 (鈪，

本文編號(hào)：1602309