铂卟啉氧传感器光学性能与生物应用研究

发布时间：2018-02-20 04:26

本文关键词： 铂卟啉氧传感器双亲高分子胶束生物　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：近年来,由于有机金属复合物光学氧传感器在检测环境中氧气浓度过程中不消耗氧气,较长的荧光寿命,操作简单,能够应用于细胞内与细胞外氧气检测等优点成为研究热点。相比于其他有机金属复合物,金属卟啉类氧传感器在室温下有着更强荧光或磷光和更高的灵敏度而被广泛应用于各种领域,如航天、环境、食品工程和医疗等。本论文以铂卟啉为高效的氧传感器,设计了两种类型的具有良好生物相容性的氧传感器材料,分别研究了它们的光学性能,对氧气的响应过程,和实时监测大肠杆菌和成骨细胞以及海拉细胞新陈代谢过程中的氧气消耗情况的应用,最后实现了细胞成像。首先,我们设计了双亲高分子来包覆四(五氟苯基)铂卟啉(Pt2+tetrakis(pentafluorophenyl)porphyrin,Pt TFPP)制作成氧传感器胶束,解决了其不能溶于水和低的荧光量子效率问题,并建立比率型的传感体系,提高了氧传感器在检测氧浓度过程中的稳定性和灵敏度。进而以Pt TFPP作为电子受体,对氧气不敏感的聚合物点作为电子给体,设计了具有荧光共振能量转移(Fluorescence Resonance Energy Transfer,FRET)效应的纳米级氧传感器,并通过荧光共振能量转移增强荧光团强度效应进一步提高氧传感器的荧光量子效率。其次,由于双亲高分子具有良好的生物相容性,我们将氧传感器实时检测了大肠杆菌(E.coli)和成骨细胞、海拉细胞在增殖过程中的氧气消耗情况,实现了将我们所设计的传感器在生物领域的应用。由于氧传感器的尺寸为纳米级别,容易被细胞内吞。通过海拉细胞的荧光成像图(Confocal images),我们可以清晰地观察到氧传感器被细胞内吞,为后续光动力治疗(PDT)实验提供有力的理论依据。最后,为了进一步扩大应用,我们将铂卟啉氧传感器修饰成为大分子单体,与其他具有良好生物相容性的单体进行共聚,合成了具有氧气敏感的双亲高分子。氧气敏感的双亲高分子直接用于溶解氧的检测,发现氧传感器能够准确测量溶解氧含量。随后,双亲高分子包覆参比探针建立了比率型的荧光传感体系提高氧传感器在检测氧浓度过程中的稳定性和灵敏度。与上一部分工作一样,我们依然将传感器应用到实时检测大肠杆菌和成骨细胞、海拉细胞在增殖过程中的氧气消耗情况。实验证明,氧传感器可以明显地分辨出不同细菌和细胞密度下氧气消耗速率的不同。
[Abstract]:In recent years, due to the fact that the organometallic complex optical oxygen sensor does not consume oxygen during the detection of oxygen concentration in the environment, it has a long fluorescence life and is easy to operate. Compared with other organometallic complexes, the advantages of being able to be applied to the detection of intracellular and extracellular oxygen have become a hot topic. Metalloporphyrin oxygen sensors have been widely used in many fields, such as aerospace, environment, food engineering and medical treatment, because of their strong fluorescence or phosphorescence and higher sensitivity at room temperature. Two kinds of oxygen sensor materials with good biocompatibility were designed and their optical properties and response to oxygen were studied. And real-time monitoring of oxygen consumption in the metabolism of Escherichia coli and osteoblasts, as well as in the metabolism of Haila cells, and finally achieved cell imaging. We have designed amphiphilic macromolecules to coat tetrakisso-pentafluorophenylphenol porphyrinPt (TFPPPN) to prepare oxygen sensor micelles, which can solve the problems of insoluble water and low fluorescence quantum efficiency, and establish a proportional sensing system. The stability and sensitivity of oxygen sensor in detecting oxygen concentration are improved, and Pt TFPP is used as electron acceptor and polymer point which is not sensitive to oxygen is used as electron donor. A nanoscale oxygen sensor with fluorescence resonance Resonance Energy transfer transfer effect is designed, and the fluorescence quantum efficiency of oxygen sensor is further enhanced by fluorescence resonance energy transfer enhanced fluorescence cluster intensity effect. Due to the good biocompatibility of amphiphilic polymers, oxygen sensors were used to detect the oxygen consumption of E. coli and osteoblasts in real time. The application of the biosensor we designed in the biological field is realized. Because the oxygen sensor is nanometer in size, It is easy to be ingested by cells. Through the fluorescence imaging of Haila cells, we can clearly observe that oxygen sensors are swallowed by cells, which provides a powerful theoretical basis for the subsequent photodynamic therapy (PDT) experiment. Finally, in order to further expand the application, We modified the platinum porphyrin oxygen sensor into macromolecular monomers and copolymerized with other monomers with good biocompatibility to synthesize oxygen sensitive amphiphilic polymers which were directly used in the detection of dissolved oxygen. It was found that the oxygen sensor could accurately measure the dissolved oxygen content. Amphiphilic polymer coated reference probe has been established to improve the stability and sensitivity of oxygen sensor in the process of detecting oxygen concentration. We still use the sensor to detect the oxygen consumption of Escherichia coli and osteoblasts in real time. The oxygen sensor can distinguish the difference of oxygen consumption rate between different bacteria and cell density.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TP212

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