【作者】 王鹏；

【导师】 胡利明；

【作者基本信息】 北京工业大学 ， 生物学， 2021， 硕士

【摘要】 荧光成像技术广泛应用于病理检测和疾病诊断。作为近红外荧光探针的花菁类荧光染料区别于传统的近红外荧光探针,在正常条件下荧光微弱,能被GSH激活产生较强荧光从而提高荧光成像性能。但是菁染料的靶向性低,能被肝脏、肾脏快速代谢清除,不利于疾病检测。针对这一问题,设计合成了两种两亲性双硫键双菁染料（SO₃Cy7-ss-Cy7）和（SO₃Cy7-ss-Cy7.5）,利用其结构本身两亲性在水溶液中自组装成纳米粒子。由于聚集淬灭,本身不发射荧光或仅有微弱荧光,当其与GSH反应后二硫键断裂导致聚集性降低从而产生强烈荧光。主要研究工作为:设计两种两亲性双硫键双菁染料并对其合成方法进行了研究,利用MODLI-TOF-MS、¹H NMR、¹³C NMR等对目标探针进行结构测定,对自组装成纳米粒子进行形貌、粒径等进行分析;利用紫外-可见光谱和荧光分光光度计进行光谱学性质的研究;对探针进行了抗干扰能力、动力学响应、pH稳定性、细胞毒性及其活细胞成像、动物成像等进行研究。结果显示:两种探针都具有良好的特异性、稳定性和灵敏度。利用MTS法测定了两种两亲性双硫键双菁荧光探针对He La细胞毒性、使用共聚焦显微镜对两种探针进行了活细胞成像以及荷瘤裸鼠体内成像。细胞水平的测试结果显示探针几乎没有毒性,活细胞荧光成像和荷瘤裸鼠体内成像清晰,该探针具有良好的肿瘤细胞靶向性,为肿瘤细胞的荧光检测提供了新的方法。更多还原

【Abstract】 Fluorescence imaging technology is widely used in pathological detection and disease diagnosis.The cyanine-based fluorescent dye as a near-infrared fluorescent probe is different from the traditional near-infrared fluorescent probe.Under normal conditions,it has weak fluorescence and can be activated by GSH to produce strong fluorescence to improve the fluorescence imaging performance.However,cyanine dyes have low targeting properties and can be rapidly metabolized and eliminated by the liver and kidneys,which is not conducive to disease detection.In response to this problem,two amphiphilic disulfide bond biscyanine dyes（SO₃Cy7-ss-Cy7）and（SO₃Cy7-ss-Cy7.5）were designed and synthesized.The structure itself was amphiphilic and self-assembled in aqueous solution.Nanoparticles.Due to aggregation quenching,it does not emit fluorescence or only weak fluorescence.When it reacts with GSH,the disulfide bond breaks,resulting in decreased aggregation and strong fluorescence.The main research work is:designing two amphiphilic disulfide bond dicyanine dyes and studying their synthesis methods,using MODLI-TOF-MS,1H NMR,13C NMR,etc.to determine the structure of the target probe,and to self-assemble The morphology,particle size,etc.of nanoparticles are analyzed;UV-visible spectroscopy and fluorescence spectrophotometer are used to study the spectroscopic properties;the probes are tested for anti-interference ability,kinetic response,pH stability,cytotoxicity and Its live cell imaging,animal imaging,etc.are studied.The results show that both probes have good specificity,stability and sensitivity.The MTS method was used to determine the toxicity of two amphiphilic disulfide bond double cyanine fluorescent probes to He La cells.The two probes were used for live cell imaging using a confocal microscope and tumor-bearing nude mice were imaged in vivo.Cell-level test results show that the probe is almost non-toxic,live cell fluorescence imaging and in vivo imaging of tumor-bearing nude mice are clear.The probe has good tumor cell targeting and provides a new method for fluorescent detection of tumor cells.更多还原