高压在线荧光检测研究
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摘要
海底热液系统及其生态系统的发现是近几十年来自然科学界最重要的发现之一。对海底热液系统中的极端环境生物和生物地球化学过程的研究工作对于揭示生命起源、生命演化以及生物成矿机制等重大科学问题具有重要意义。高压物理学作为一门重要的交叉学科已广泛应用于生物科学研究中,特别是极端环境生物的发现,使高压物理学成为极端生物及各类生物分子研究的重要工具之一。同时随着检测技术的进步,光谱检测法的引入使高压在线检测研究成为可能。其中荧光光谱法的引入使高压在线检测更加灵敏可靠,有力的推动了极端环境生物分子的研究,包括生物分子间的相互关系,生物分子的结构变化乃至对极端生物生命活动、适应机制等。
本论文首先对高压在线检测的国内外现状进行了总结和分析,建立了新的可用于普通荧光分光光度计的小型高压在线荧光检测装置。该装置不但可以实时在线荧光检测,还可以与大型高压装置耦合以进行流动体系的高压在线荧光检测。以荧光物质虎红为研究对象,对该装置进行了试验,在此基础研究了虎红的高压荧光特性。实验结果表明该装置可以很好的用于高压在线荧光研究,同时首次揭示了这一物质的高压荧光特性。
其次,色氨酸是多种海洋生物大分子的内源荧光物质之一,色氨酸的高压荧光检测无论在海洋生物总量的测定还是在生物大分子结构研究方面都具有重要的意义。据此,本文系统地考察了压力,金属离子,pH值等因素对色氨酸荧光效应的影响。获得了色氨酸在高压条件下新的荧光特性数据,系统化的阐述了压力对荧光效应的影响。
另外,针对深海极端环境条件下生物研究中长期实时观测的困难,建立了新的可用于实验室在线显微观测和培养的高压显微观测装置。该装置不但可以实现实验室高压显微观测,还可以对生物进行实验室培养。利用该装置,实现了浮游生物的高压实时在线观测。
论文主要包括以下几个部分：第一部分综述了目前高压检测研究的现状,分析了目前存在的问题及发展方向；第二部分对高压在线荧光检测基础理论进行分析和总结；第三部分介绍了自行研制开发的高压在线荧光检测装置以及虎红高压荧光检测；第四部分介绍了色氨酸的相关高压荧光检测实验,并系统化的阐述了压力对荧光效应的影响。第五部分介绍了自行研制的高压显微观测培养装置并进行了浮游生物的在线高压显微观测。最后一部分对本论文进行了总结并对今后相关研究进行了展望。
As an important parameter, pressure has been widely used in biology research. Especially after spectacular biological communities were discovered in extreme environments such as hydrothermal vent, pressure has attracted more attentions in the research of extremophiles and biomolecule. As the use of spectroscopy method in high pressure research, the on-line detection under high pressure becames possible. Especially fluorescence spectrometry was used in high pressure research, the experiment becames more sensitive and reliable. At present, There is tremendous progress in the researches about biomolecule under pressures. These researches are very important in studing the interaction of biomolecular, the changes of biomolecular structure, and the activities of extremophiles et al. With the progress of spectrum methods under pressures, the devices of on-line detection under high pressure were developed.
First, the paper summarized the researches about on-line fluorescence detecion. Then We developed an on-line fluorescence detection device which could be installed in common fluorescence spectrophotometer. This device could be used for on-line fluorescence detection under high pressure, and also could be connected with a large-scale device for flow-system detection on-line. With this system, the fluorescence spectrum of Rose Bengal under pressure was studied. The results showed that the device was valid for on-line fluorescence detection under high pressure. At the same time, the results of Rose Bengal provied important direct evidence for biomolecule researches.
Second, tryptophan is an important intrinsic fluorophore in reasearches of the deep sea, so we studied the fluorescence spectrum of pure tryptophan and metal ions/tryptophan mixtures under high pressures. The results showed that the effect of pressure on pure tryptophan and metal ions/tyrptophan mixtures must be considered in researches of macromolecular under pressure.
Another important section of this paper was a high pressure on-line observation system for microorganism. The system could be used with fluorescence microscope or long distance microscope. Using this system, we studied Cyclops under different pressures. This was important in researches of the distribution of plankton and activities under high pressures.
本论文首先对高压在线检测的国内外现状进行了总结和分析,建立了新的可用于普通荧光分光光度计的小型高压在线荧光检测装置。该装置不但可以实时在线荧光检测,还可以与大型高压装置耦合以进行流动体系的高压在线荧光检测。以荧光物质虎红为研究对象,对该装置进行了试验,在此基础研究了虎红的高压荧光特性。实验结果表明该装置可以很好的用于高压在线荧光研究,同时首次揭示了这一物质的高压荧光特性。
其次,色氨酸是多种海洋生物大分子的内源荧光物质之一,色氨酸的高压荧光检测无论在海洋生物总量的测定还是在生物大分子结构研究方面都具有重要的意义。据此,本文系统地考察了压力,金属离子,pH值等因素对色氨酸荧光效应的影响。获得了色氨酸在高压条件下新的荧光特性数据,系统化的阐述了压力对荧光效应的影响。
另外,针对深海极端环境条件下生物研究中长期实时观测的困难,建立了新的可用于实验室在线显微观测和培养的高压显微观测装置。该装置不但可以实现实验室高压显微观测,还可以对生物进行实验室培养。利用该装置,实现了浮游生物的高压实时在线观测。
论文主要包括以下几个部分：第一部分综述了目前高压检测研究的现状,分析了目前存在的问题及发展方向；第二部分对高压在线荧光检测基础理论进行分析和总结；第三部分介绍了自行研制开发的高压在线荧光检测装置以及虎红高压荧光检测；第四部分介绍了色氨酸的相关高压荧光检测实验,并系统化的阐述了压力对荧光效应的影响。第五部分介绍了自行研制的高压显微观测培养装置并进行了浮游生物的在线高压显微观测。最后一部分对本论文进行了总结并对今后相关研究进行了展望。
As an important parameter, pressure has been widely used in biology research. Especially after spectacular biological communities were discovered in extreme environments such as hydrothermal vent, pressure has attracted more attentions in the research of extremophiles and biomolecule. As the use of spectroscopy method in high pressure research, the on-line detection under high pressure becames possible. Especially fluorescence spectrometry was used in high pressure research, the experiment becames more sensitive and reliable. At present, There is tremendous progress in the researches about biomolecule under pressures. These researches are very important in studing the interaction of biomolecular, the changes of biomolecular structure, and the activities of extremophiles et al. With the progress of spectrum methods under pressures, the devices of on-line detection under high pressure were developed.
First, the paper summarized the researches about on-line fluorescence detecion. Then We developed an on-line fluorescence detection device which could be installed in common fluorescence spectrophotometer. This device could be used for on-line fluorescence detection under high pressure, and also could be connected with a large-scale device for flow-system detection on-line. With this system, the fluorescence spectrum of Rose Bengal under pressure was studied. The results showed that the device was valid for on-line fluorescence detection under high pressure. At the same time, the results of Rose Bengal provied important direct evidence for biomolecule researches.
Second, tryptophan is an important intrinsic fluorophore in reasearches of the deep sea, so we studied the fluorescence spectrum of pure tryptophan and metal ions/tryptophan mixtures under high pressures. The results showed that the effect of pressure on pure tryptophan and metal ions/tyrptophan mixtures must be considered in researches of macromolecular under pressure.
Another important section of this paper was a high pressure on-line observation system for microorganism. The system could be used with fluorescence microscope or long distance microscope. Using this system, we studied Cyclops under different pressures. This was important in researches of the distribution of plankton and activities under high pressures.
引文
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