有机小分子探针的设计合成及其在生物成像上的应用

发布时间：2018-03-31 12:02

本文选题：荧光探针　切入点：双光子　出处：《济南大学》2017年硕士论文

【摘要】：近年来,有机小分子荧光探针,作为一种高灵敏度、高选择性、低成本、免损伤的检测手段,能够对靶标物实现原位、实时、可视化的检测,尤其是在生物成像方面的应用,被世界各地的课题组广泛应用于各种类型的靶标物分子的示踪标记、生理过程检测和临床医学分析等方面。氟离子,作为电负性最强,离子半径最小的阴离子,是生物体内不可缺少的微量元素,具有十分重要的生理功能。氟元素是构成骨骼,牙齿的必备元素,人体内缺少氟元素可能会导致骨质疏松,龋齿等疾病;氟元素过量,可能会引起氟骨病,氟斑牙等疾病。鉴于此,对氟离子的检测,具有十分重要的意义。过氧化氢,活性氧物种之一,通过细胞因子,神经递质等刺激细胞NADPH(还原型辅酶)氧化酶复合物活化而产生,对免疫过程,细胞信号传递、细胞衰老、凋亡等生理过程具有十分重要的作用,近年来的研究发现,过氧化氢含量异常,与癌症、糖尿病、神经变性疾病和心脑血管等疾病具有十分密切的联系。双光子荧光探针具有背景干扰低、穿透能力强等优势,比单光子探针更适合应用于活细胞以及组织成像。基于此,论文的第一部分,本课题组以叔丁基二甲基氯硅烷为识别基团设计合成了一种双光子氟离子荧光探针,荧光增强倍数接近10倍,该探针对氟离子具有选择性好,灵敏度高,检测限低,细胞毒性低,生物相容性好,水溶性好等优势,经计算,该探针对氟离子的检测限低至7.8×10-7 M。该探针分子已经成功应用于人宫颈癌细胞(HeLa)单、双光子成像,并将该氟离子荧光探针应用于小鼠组织成像。论文的第二部分,本课题组还利用4-硼酯溴苄作为识别基团设计合成了一种双光子过氧化氢探针,荧光增强倍数约为40倍,该探针能够专一识别活性氧物种中的过氧化氢,响应速度快,灵敏度高,检测限低。经测试计算,其检测限达1.46×10-7M。并将该探针成功应用于HeLa单、双光子外源性成像,小鼠巨噬细胞(RAW 264.7)单、双光子内源性成像,以及小鼠肝脏组织单、双光子成像。论文的第三部分,我们采用荧光量子产率高,且stokes位移大的菲咪唑荧光团,设计合成了两种菲咪唑荧光团的过氧化氢探针,此类探针响应迅速,灵敏度高,检测限低。其对过氧化氢的检测限分别达到了1.48×10-7 M、1.08×10-7 M。并将该类探针成功应用于HeLa细胞的内源性、外源性细胞成像,OGD/R(氧/葡萄糖灌注实验)以及小鼠活体成像。
[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.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R318;O657.3

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