FUNDAMENTAL STUDY AND APPLICATION OF OPTICAL CHROMATOGRAPHY - TRACE ANALYSIS OF BIOACTIVE SUBSTANCES AND PRECISE DETERMINATION OF MICROORGANISM'S POWER

提出了一种利用光辐射压力进行微粒分离的新方法--光色谱，并将其应用于蛋白质、微生物能力和红细胞变形性的测定。为了评价光学层析的性能，利用射线光学模型从理论上推导了一些方程。根据理论计算结果，可以通过增大激光功率和降低流量来扩展动态范围。将光学层析技术应用于蛋白质的痕量分析。两个涂有抗体的聚苯乙烯小球在抗原存在的情况下结合在一起。结合珠(B)和游离珠(F)可通过光色谱分离，其B/F值与抗原(蛋白质)的浓度相关。正向和反向免疫反应的速率分别通过测量免疫微球的形成时间和解离时间来独立确定。这项被称为光学漏斗的技术利用了一种系统，在该系统中，微生物悬浮在流动的介质中，辐射压力以相反的方向施加。在平衡状态下，微生物被捕获。在这一点上，它有能力依靠自己的力量逃脱。使用这项技术，有可能测量微生物的力量的大小。光学漏斗还用于测定牛精子细胞的活动力。测定了不同pH值和不同钾离子浓度的水溶液中精子细胞的活动力。它有可能在三个小时内测量超过250个精子细胞。用近红外激光光学漏斗评价红细胞的变形性。测得年轻细胞的红细胞伸长率为2.4±0.6，老年细胞的伸长率为2.1±0.5，与另一种方法测得的结果一致。

A new method for separation of particles by optical radiation pressure, named as optical chromatography, was developed and applied to the determination of protein, microorganism's power, and deformability of erythrocyte. In order to evaluate the performance of optical chromatography, a number of equations are theoretically derived using a ray-optics model. Based on the results obtained by theoretical calculation, the dynamic range can be extended by increasing laser power and by decreasing flow rate. Optical chromatography was applied to trace analysis of protein. Two polystyrene beads, coated with antibody, are combined in the presence of an antigen. The bound (B) and free (F) beads were readily separated by optical chromatography, and the B/F ratio could be correlated with the concentration of antigen (protein). The rates of the forward and reverse immunological reactions were independently determined by measuring the time of formation and dissociation, respectively, of the immunobeads. The technique, called as optical funnel, utilized a system in which a microorganism is suspended in a flowing medium and radiation pressure, which is applied in the opposite direction. At equilibrium, the microorganism is captured. At this point, it has the capability to escape on its own power. Using this technique, it is possible to measure the magnitude of the microorganism's power. The optical funnel was also applied to the determination of the motility force of bovine sperm cells. The motility force of the sperm cell was measured in the aqueous solution at different pH values and potassium ion concentrations. It was possible to measure more than 250 sperm cells in three hours. Deformability of erythrocyte was evaluated by optical funnel using near infrared laser. Elongation of erythrocyte was measured as 2.4 ± 0.6 for young cells and 2.1 ± 0.5 for old cells, which were in good agreement with the values obtained by the other method.