【摘要】：近年來,熒光檢測法因其具有操作簡便、靈敏度高、實時快速分析等優(yōu)點,廣泛應(yīng)用于環(huán)境檢測分析和生物成像等領(lǐng)域。其中,具有良好水溶性的多功能熒光探針能夠高選擇性檢測復(fù)雜體系中兩種或以上分析物已成為研究熱點。因此,本論文中我們設(shè)計合成了三種新穎的水溶性雙功能熒光探針,分別實現(xiàn)了對銅離子(Cu~(2+))和硫離子(S~(2-))、四氫呋喃中水含量和pH值、次氯酸根(ClO~-)和溫度的快速、靈敏、選擇性的熒光檢測。本論文的主要內(nèi)容和結(jié)果如下:1.基于熒光能量共振轉(zhuǎn)移(FRET)原理,我們將異硫氰酸熒光素(FITC)共價鍵連接到支化聚乙烯亞胺(b-PEI)分子上,制得了一種水溶性雙功能熒光聚合探針(PF)。PF能夠選擇性地絡(luò)合Cu~(2+)生成PF-Cu~(2+)絡(luò)合物,PF體系熒光減弱,這是由于熒光素(給體)和PF-Cu~(2+)絡(luò)合物(受體)之間發(fā)生了有效地FRET。當(dāng)向上述PF-Cu~(2+)體系中加入S~(2-),S~(2-)和Cu~(2+)結(jié)合形成穩(wěn)定的CuS沉淀物,PF體系熒光恢復(fù),實現(xiàn)了對Cu~(2+)和S~(2-)高選擇性檢測。此外,該探針PF被成功應(yīng)用于自來水和人血清中Cu~(2+)和S~(2-)的高靈敏度熒光檢測。2.以對苯二胺為原料,采用水熱法,我們合成了一種水溶性雙功能紅色熒光碳量子點(R-Cdots)。隨著pH的增大(2.0~9.0),R-Cdots在610 nm處的熒光逐漸增大,并且在5.0~9.0之間,熒光強(qiáng)度值和pH有較好的線性關(guān)系,表明R-Cdots能夠靈敏地檢測復(fù)雜生物樣品的pH。此外,R-Cdots具有較強(qiáng)的溶劑極性依賴性,其熒光發(fā)射波長隨溶劑極性增大而發(fā)生紅移(~80 nm)。隨著四氫呋喃中的水含量增加,其熒光發(fā)射波長發(fā)生紅移且熒光強(qiáng)度減弱(綠色變?yōu)榧t色),檢測限低至0.04%(體積分?jǐn)?shù)),實現(xiàn)了對四氫呋喃中水含量的熒光定量檢測。3.我們將異硫氰酸羅丹明B通過共價鍵的方式連接到熒光硅納米粒子表面氨基上,制得一種水溶性雙功能比率型熒光納米探針(DRFS)。當(dāng)向DRFS溶液中加入ClO~-,硅納米粒子的熒光(445 nm)減弱,而羅丹明B的熒光(576 nm)基本保持不變,從而實現(xiàn)了對ClO~-高選擇性熒光檢測。DRFS不僅可以測定人血清中ClO~-的濃度,而且還能夠?qū)?xì)胞中外源性和內(nèi)源性的ClO~-進(jìn)行熒光成像。另外,DRFS也可作為一種溫度熒光傳感器。隨著溫度的升高,羅丹明B的熒光(576 nm)逐漸下降,而硅納米粒子的熒光(445 nm)基本保持不變,并且在20℃~60℃之間,熒光強(qiáng)度比值(I576/I445)和溫度有較好的線性關(guān)系,表明DRFS能夠定量監(jiān)測生物體系中的溫度變化。
[Abstract]:In recent years, fluorescence detection method has been widely used in the field of environmental detection and biological imaging because of its advantages of simple operation, high sensitivity, real-time rapid analysis and so on. Among them, multifunctional fluorescent probes with good water solubility can detect two or more analytes in complex systems with high selectivity. Therefore, we have designed and synthesized three novel water-soluble bifunctional fluorescent probes in this thesis. We have achieved the sensitivity and rapidity of water content and pH value, hypochlorite (ClO~-) and temperature of copper ion (Cu~ (2) and sulfur ion (S ~ (2), tetrahydrofuran), respectively. Selective fluorescence detection. The main contents and results of this thesis are as follows: 1. Based on the principle of fluorescence energy resonance transfer (FRET), we connect the covalent bond of fluorescein isothiocyanate (FITC) to the branched polyvinyleneimine (b-PEI) molecule. A water-soluble bifunctional fluorescent polymerization probe (PF) 路PF was prepared, which can selectively complexate Cu~ (2) to form PF-Cu~ (2) complex. The fluorescence of PF-Cu~ (2) complex / PF system is weakened, which is due to the effective FRET. between fluorescein (donor) and PF-Cu~ (2) complex (receptor). When S2- and Cu~ (2) were added to the PF-Cu~ (2) system to form a stable CuS precipitate, the fluorescence recovery was achieved, and the high selectivity detection of Cu~ (2) and S2- was realized. In addition, the probe PF was successfully applied to the detection of Cu~ (2) and S2- in tap water and human serum. A water-soluble bifunctional red fluorescent carbon quantum dot (R-Cdots) was synthesized from p-phenylenediamine by hydrothermal method. With the increase of pH (2.0 ~ 9.0), the fluorescence of R-Cdots at 610 nm increases gradually, and the fluorescence intensity has a good linear relationship with pH in the range of 5.0 ~ 9.0, which indicates that R-Cdots can detect pH. of complex biological samples sensitively. In addition, R-Cdots have strong solvent polarity dependence, and their fluorescence emission wavelengths shift red with the increase of solvent polarity (80 nm). With the increase of water content in tetrahydrofuran, the fluorescence emission wavelength shifts red and the fluorescence intensity decreases (green to red), the detection limit is as low as 0.04% (volume fraction), and the fluorescence quantitative detection of water content in tetrahydrofuran is realized. We have connected Rhodamine isothiocyanate B to the surface amino of fluorescent silicon nanoparticles by covalent bonding to prepare a water-soluble bifunctional fluorescent nano-probe (DRFS). When the fluorescence (445 nm) of ClO~-, silicon nanoparticles was weakened in DRFS solution, but the fluorescence of Rhodamine B (576 nm) remained unchanged, the highly selective fluorescence detection of ClO~- could not only determine the concentration of ClO~- in human serum. Moreover, fluorescent imaging of exogenous and endogenous ClO~- in cells is also possible. In addition, DRFS can also be used as a temperature fluorescence sensor. With the increase of temperature, the fluorescence of rhodamine B (576 nm) decreased gradually, while the fluorescence of silicon nanoparticles (445 nm) remained unchanged, and the fluorescence intensity ratio (I576/I445) had a good linear relationship with temperature between 20 鈩，

本文編號：2258002