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针对离线单微粒跟踪缺乏运动观测实时性、无法对单微粒运动状态进行快速反馈等问题,设计出一套能实时进行单微粒跟踪的系统。该系统以荧光单微粒成像系统为硬件基础,以LabVIEW为软件开发平台,将CCD荧光图像采集、荧光显微镜操作和单微粒跟踪运算融为一体,实现荧光图像采集与单微粒跟踪的并行执行和实时控制。在普通个人计算机上利用该系统对100帧连续随机运动的单微粒荧光图像的仿真和并六苯分子在云母基体的扩散实验研究表明,在运算图像高达百万级像素时,该系统可实现4~5帧/秒的跟踪速率。这对荧光标记大分子和有机体的扩散认知、分子马达运动特性研究和荧光有机分子薄膜生长机理解析等具有重要的意义。
In view of the lack of real-time tracking of off-line single-particle tracking and the inability to provide fast feedback on the movement of single-particle, a system of tracking single-particle in real-time is designed. The system takes the fluorescent single particle imaging system as the hardware base and LabVIEW as the software development platform, which integrates the CCD fluorescence image acquisition, the fluorescence microscope operation and the single particle tracking operation to realize the parallel execution of fluorescent image acquisition and single particle tracking and real-time control. The simulation of a single-particle fluorescence image of 100 frames continuous random motion and the diffusion experiment of hexa-benzene molecules on the mica matrix are carried out on an ordinary personal computer. The experimental results show that when the image is up to millions of pixels, the system can achieve 4 ~ 5 frames per second tracking rate. This is of great significance for the diffusion recognition of fluorescently labeled macromolecules and organisms, kinetic characterization of molecular motors and the analysis of the growth mechanism of fluorescent organic molecules.