本文選題：熒光探針　切入點：雙光子　出處：《濟南大學(xué)》2017年碩士論文

【摘要】：近年來,有機小分子熒光探針,作為一種高靈敏度、高選擇性、低成本、免損傷的檢測手段,能夠?qū)Π袠?biāo)物實現(xiàn)原位、實時、可視化的檢測,尤其是在生物成像方面的應(yīng)用,被世界各地的課題組廣泛應(yīng)用于各種類型的靶標(biāo)物分子的示蹤標(biāo)記、生理過程檢測和臨床醫(yī)學(xué)分析等方面。氟離子,作為電負(fù)性最強,離子半徑最小的陰離子,是生物體內(nèi)不可缺少的微量元素,具有十分重要的生理功能。氟元素是構(gòu)成骨骼,牙齒的必備元素,人體內(nèi)缺少氟元素可能會導(dǎo)致骨質(zhì)疏松,齲齒等疾病;氟元素過量,可能會引起氟骨病,氟斑牙等疾病。鑒于此,對氟離子的檢測,具有十分重要的意義。過氧化氫,活性氧物種之一,通過細(xì)胞因子,神經(jīng)遞質(zhì)等刺激細(xì)胞NADPH(還原型輔酶)氧化酶復(fù)合物活化而產(chǎn)生,對免疫過程,細(xì)胞信號傳遞、細(xì)胞衰老、凋亡等生理過程具有十分重要的作用,近年來的研究發(fā)現(xiàn),過氧化氫含量異常,與癌癥、糖尿病、神經(jīng)變性疾病和心腦血管等疾病具有十分密切的聯(lián)系。雙光子熒光探針具有背景干擾低、穿透能力強等優(yōu)勢,比單光子探針更適合應(yīng)用于活細(xì)胞以及組織成像�；诖�,論文的第一部分,本課題組以叔丁基二甲基氯硅烷為識別基團設(shè)計合成了一種雙光子氟離子熒光探針,熒光增強倍數(shù)接近10倍,該探針對氟離子具有選擇性好,靈敏度高,檢測限低,細(xì)胞毒性低,生物相容性好,水溶性好等優(yōu)勢,經(jīng)計算,該探針對氟離子的檢測限低至7.8×10-7 M。該探針分子已經(jīng)成功應(yīng)用于人宮頸癌細(xì)胞(HeLa)單、雙光子成像,并將該氟離子熒光探針應(yīng)用于小鼠組織成像。論文的第二部分,本課題組還利用4-硼酯溴芐作為識別基團設(shè)計合成了一種雙光子過氧化氫探針,熒光增強倍數(shù)約為40倍,該探針能夠?qū)Ｒ蛔R別活性氧物種中的過氧化氫,響應(yīng)速度快,靈敏度高,檢測限低。經(jīng)測試計算,其檢測限達1.46×10-7M。并將該探針成功應(yīng)用于HeLa單、雙光子外源性成像,小鼠巨噬細(xì)胞(RAW 264.7)單、雙光子內(nèi)源性成像,以及小鼠肝臟組織單、雙光子成像。論文的第三部分,我們采用熒光量子產(chǎn)率高,且stokes位移大的菲咪唑熒光團,設(shè)計合成了兩種菲咪唑熒光團的過氧化氫探針,此類探針響應(yīng)迅速,靈敏度高,檢測限低。其對過氧化氫的檢測限分別達到了1.48×10-7 M、1.08×10-7 M。并將該類探針成功應(yīng)用于HeLa細(xì)胞的內(nèi)源性、外源性細(xì)胞成像,OGD/R(氧/葡萄糖灌注實驗)以及小鼠活體成像。
[Abstract]:In recent years, organic small molecular fluorescence probes, as a high sensitivity, high selectivity, low cost, non-damage detection method, can be used for in-situ, real-time and visual detection of target, especially in the field of biological imaging. It has been widely used in various kinds of target molecular tracer labeling, physiological process detection and clinical medical analysis, etc. Fluorine ion is the most electronegativity anion with the smallest ion radius. Fluorine is an indispensable trace element in the body and has very important physiological functions. Fluorine is an essential element in the formation of bone and teeth. The lack of fluoride in the human body may lead to osteoporosis, dental caries and other diseases. May cause fluorosis, dental fluorosis and so on. In view of this, the detection of fluorine ions has very important significance. Hydrogen peroxide, one of the reactive oxygen species, through cytokines, Neurotransmitter stimulates the activation of NADPH oxidase complex, which plays an important role in immune process, cell signal transmission, cell senescence, apoptosis and other physiological processes. The abnormal content of hydrogen peroxide is closely related to cancer, diabetes, neurodegenerative diseases and cardiovascular and cerebrovascular diseases. The single photon probe is more suitable for living cell and tissue imaging. In the first part of this paper, we designed and synthesized a two-photon fluorine fluorescence probe using tert-#china_person0# dimethyl chlorosilane as the recognition group. The fluorescence enhancement multiple is close to 10 times. The probe has the advantages of good selectivity, high sensitivity, low detection limit, low cytotoxicity, good biocompatibility, good water solubility and so on. The detection limit of fluorine ion is as low as 7.8 脳 10 ~ (-7) M. the probe molecule has been successfully applied to single and two-photon imaging of human cervical cancer cell line HeLa, and the fluorine fluorescence probe has been applied to tissue imaging of mice. We also designed and synthesized a two-photon hydrogen peroxide probe using 4-boron ester bromobenzyl bromide as the recognition group. The fluorescence enhancement is about 40 times. The probe can specifically identify hydrogen peroxide in active oxygen species and the response speed is fast. The detection limit was 1.46 脳 10 ~ (-7) M.The probe was successfully used in HeLa single, two-photon exogenous imaging, mouse macrophage RAW264.7) single, two-photon endogenous imaging, and mouse liver tissue monolayer. Two-photon imaging. In the third part of the thesis, we designed and synthesized two kinds of hydrogen peroxide probes with high fluorescence quantum yield and large stokes shift. The detection limit of hydrogen peroxide was 1.48 脳 10 ~ (-7) MN 1.08 脳 10 ~ (-7) M.The probe was successfully applied to endogenous HeLa cells, exogenous cell imaging (OGD / R) and in vivo imaging of mice.
【學(xué)位授予單位】：濟南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R318;O657.3

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