脉冲升温时间分辨红外光谱仪的分析-生物物理学专业毕业论文.docxVIP

摘要摘要． 摘要 摘要． 蛋白质折叠是当前生命科学的重大问题，因为它关乎生命的正常运转。我们 实验室搭建激光脉冲升温实验技术平台，用红外激光脉冲使样品升温进而诱发蛋 白质开折叠，以时间分辨的红外光谱作为检测手段跟踪其动力学，实验室的前人 们已经成功的解释了重水作为溶剂的很多蛋白包括二硫键异构酶DsbC和热休克 蛋白DegP，光敏黄蛋白PYP等的热诱导的开折叠过程。脉冲升温时间分辨红外 光谱研究蛋白质折叠动力学过程，直接获得的信息是吸光度的变化，所以探测光 和泵浦光的时空稳定性也就成了影响测量精度的主要因素，实验室现行的泵浦光 和探测光系统都具有不同程度的误差，这样我们就必须采取双光路绞基线的方式 来消除这种误差，又带来了新的系统误差。 本文着眼于提高脉冲升温时间分辨红外光谱的精度，对泵浦光系统和探测光 系统进行优化，重点阐述光路及元器件的搭建和优化，将拉曼频移池系统进行优 化，使出射单脉冲能量提高至84．8mJ，提高了近36％，并利用光声探测器对单脉 冲能量进行时间稳定性分析，稳定性可达到3％。接着对脉冲光斑进行了分析优化， 光斑由椭圆型变成了圆形，能量分布也更均匀了。另外，对实验室的钬激光器进 行了升级，使得单脉冲宽度从80ns缩短至40ns，使钬激光在激光脉冲升温领域 成为新的更加可靠的手段，结论：对泵浦光路和探测光路系统的优化使得脉冲升 温时间分辨红外手段在蛋白质开折叠动力学研究中成为更加可靠的手段。 关键词： 激光脉冲升温蛋白质折叠受激拉曼散射钬激光 AbstractAbstract Abstract Abstract Folding procedure of the protein has been the highlight of the biological science for a long time，because in every possible way it’S essential to all the activities of living organism．Laser pulse-induced temperature-jump(T-jump)，a technique that was very popular and useful，has been applied in our lab to trigger the unfolding of certain protein．What’s more，time—resolved infrared(IR)provide a good way to probe the subsequent unfolding kinetics and the structure ordering all through the unfolding procedure．The previous workers in this lab had successfully investigated the thermal-induced unfolding process of DsbC DegP and PYP dissolved in I)20．In the experiment，the uncertainty Was mainly brought by the time and spatial unstability of both pump and probe lights．Double light pathes and baseline matching method had to be concerned． To minimize the unstability of the whole system，optimization of both pump light and probe system is essential，Stimulated Raman Scattering(SRS)set，which Was used to trigger the unfolding kinetics，has been upgraded，and energy of single laser pulse has been raised to 84．8mJ，about 1 36 percent of the old one，and then the corresponding stability of output single laser pulse has been carefully evaluated，of course，a very smart apparatus，photon-sound laser detector invented by one of t