【作者】 金磊；

【导师】 胡晓东；

【作者基本信息】 天津大学 ， 仪器科学与技术， 2018， 硕士

【摘要】 高通量单分子力谱测量技术是单分子动力学等生物动态过程的重要研究工具之一。通过实现同时测量数十甚至上百的单分子力谱,很大程度上提高了实验效率和可靠性。然而因为需要计算的数据量十分庞大,所以缺乏相匹配的高精度实时计算方法。数字全息显微技术具有非接触、高精度及实时性等特点,是近三十年来光学微纳米检测领域的研究热点之一。本文将数字全息技术与高通量单分子力谱测量技术进行了融合,提出数字全息离心力显微测量系统,实现了对单分子动力学特征的高通量高精度实时检测。本论文研究内容和成果包括:1.系统地阐述了应用同轴数字全息显微成像技术测量微球三维位置的方法,包括数字全息测量的光路系统、基本测量原理、不同的实现方法等。并验证了将其应用于高通量单分子测量技术的可行性。2.在离心力显微镜的基础上进行改进,搭建了一种更可靠的高通量单分子力谱测量系统——数字全息离心力显微系统。阐述内容包括系统原理、光路搭建、测量理论及方法等,并对机械形变、样品池形变及粒子布朗运动等因素所造成的误差进行了分析和仿真。3.阐述了高通量单分子样品的制作方法。分析了目前高通量单分子测量样品制作方法中所存在的问题,并对基底的粘滞阻尼和特异性结合等问题进行了进一步实验研究。并提出了新的样品制作方法。实验结果数据统计后证明,改进后样品制作方法将高通量分子力谱测量效率提高了15.2倍。4.采用新方法制作的样品在数字全息离心力显微系统上进行了纳米台阶和高通量单分子拉伸实验。实验显示,该系统轴向分辨率可达到5nm,横向分辨率可达1nm。其单分子拉伸力谱曲线与目前世界公认的蠕虫曲线模型(WLC)相契合,从而同时验证了数字全息离心力显微系统以及样品制作的有效性。更多还原

【Abstract】 High-throughput single-molecule force spectroscopy(H-SMFS)is one of the most important tools for abundant biodynamic such as single-molecule dynamics.The experimental efficiency and reliability are improved via measuring tens even hundreds of single-molecules simultaneously.However,there is no matching high-precision and real-time algorithm for its mass data.Digital holographic microscopy(DHM),as one of the hotpots in the field of optical micro-nanometer detection in the past 30 years,has the characteristics of contactless,high accuracy and real-time.In this paper,DHM and H-SMFS are combined to develop a digital holographic centrifugal force microscopy(DH-CFM)system,which realizes high-throughput and high-precision detection of single molecules in real time directly.The main research contains,1.Analyzing the method of tracing the beads in three-dimensions by DHM systematically,including the optical path,principles,and different implementation methods.And it is verified that the feasibility of applying it to H-SMFS systems.2.Based on the DHM and CFM,DH-CFM system is proposed.The contents include system principles,optical path’s building,measurement theories and methods.The errors caused by mechanical deformation,sample cell deformation and particle Brownian motion are analyzed and simulated.3.The specimen preparing method for SMFS systems is described.Analyzing the problems arising from the current SMFS specimen and the relevant researches such as viscous damping and specific linkage.Then,we demonstrate a new method of specimen preparation.Experimental results show that the proposed method improves the efficiency of H-SMFS more than 15.2 times.4.The step response and DNA stretching procession have detected following the proposed method by DH-CFM.The results manifest the resolution is within 5nm and DNA extension curves accord with Worm-like-chain(WLC)which was accepted worldly.It verifies the validity both the DH-CFM system and the specimen preparing method.更多还原