新型细胞微环境检测荧光探针的设计、合成和应用

发布时间：2018-03-11 08:07

本文选题：pH值　切入点：比色/荧光　出处：《上海师范大学》2017年硕士论文　论文类型：学位论文

【摘要】：基于半菁染料,荧光碳点为荧光母体,设计、制备了三种高选择性荧光探针。利用分子内电荷转移效应(ICT),细胞内吞和靶向基团实现了环境中和和活细胞内微环境的定性、定量检测。(1)由于传统的p H计不能很好地应用在微环境如亚细胞器和复杂生物体系中的p H值动态和精确定量检测。因此迫切需要研究可监测复杂多变生物体系中p H值变化且具有高灵敏度的p H值响应分子探针。基于半菁染料合成了一种比色和荧光双模态检测p H的比率型分子探针CPH。CPH的具有合适的p Ka(6.44),使其可以聚集在溶酶体内成为靶向溶酶体的探针。CPH可监测经抗疟疾药氯喹刺激He La细胞后溶酶体内p H值的变化。将CPH固载在试纸上,可实现对环境中酸性和碱性蒸汽的可逆、动态、裸眼检测。这一概念验证展现出CPH作为检测溶酶体内p H变化的分子工具的潜在应用,同时提供了一种经济、可重复和快速对酸碱蒸汽的光学p H探针。(2)溶酶体是酸性囊状结构的细胞器,其膜内含有大量酶和蛋白质,p H值是4.5-5.5。因此,发展通过可视化荧光信号用于溶酶体标记和成像对理解与溶酶体功能紊乱相关的病理学显得尤为重要。首次设计、制备了基于荧光碳点用于标记溶酶体的纳米探针CDs-PEI-ML。该探针具有良好的水溶性和光稳定性。我们推测其成像原理是该纳米探针先通过内吞进入He La细胞,再通过吗啉基团使其聚集在溶酶体内。(3)基于半菁骨架,我们发展了一种比率型溶酶体探针CPY。该探针具有优良的溶酶体靶向性。利用该探针揭示出溶酶体的p H会随着热刺激而升高,且这个过程在短时间内是不可逆的。
[Abstract]:Three highly selective fluorescent probes were designed and prepared based on hemicyanine dyes with fluorescence carbon spots as the fluorescent parent. The intracellular endocytosis and targeting groups were used to characterize the environmental and living microenvironments by using intramolecular charge transfer effect (ICTI). (1) since the traditional pH meter can not be well applied to dynamic and accurate quantitative detection of pH values in microenvironments such as suborganelles and complex biological systems, there is an urgent need to study and monitor complex and variable biological systems. Middle pH value varied and highly sensitive molecular probe with pH response. Based on hemicyanine dye, a ratio molecular probe CPH.CPH with colorimetric and fluorescence bimodal detection of pH was synthesized, which can be aggregated. The lysosomal probe. CPH can monitor the change of pH value in helium La cells stimulated by chloroquine. The CPH was immobilized on the test paper. This proof of concept demonstrates the potential application of CPH as a molecular tool for detecting changes in pH in lysosomes and provides an economy. The lysosome is an acidic cystosomal organelle containing a large number of enzymes and proteins (pH = 4.5-5.5). The development of visual fluorescent signals for lysosomal labeling and imaging is particularly important in understanding the pathology associated with lysosomal dysfunction. A fluorescent carbon dot based probe CDs-PEI-MLwas prepared for labelling lysosomes. The probe has good water-solubility and photostability. We speculate that the imaging principle of the probe is that the probe first enters He-La cells through endocytosis. Then the morpholine group is used to aggregate it in lysosome. 3) based on hemicyanine skeleton, We have developed a ratio lysosomal probe CPY. the probe has excellent lysosomal targeting. It is revealed that the pH of lysosome increases with heat stimulation and the process is irreversible in a short time.
【学位授予单位】：上海师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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