【摘要】 利用TIRF显微镜动态实时地观测了GLUT4囊泡在脂肪细胞内的运动,并针对TIRF显微镜消散场成像的特点,提出了一种新的三维单粒子跟踪算法。算法首先针对囊泡的荧光淬灭问题,对荧光强度作了校正;然后,应用自适应的背景减除算法,分割出了运动的囊泡;最后,在跟踪过程中引入了Kalman滤波,减小了搜索的范围,从而避免了背景噪声以及虚假目标的干扰。对囊泡在胞内的长距离运动作跟踪实验,结果表明算法不但在成像平面上取得了很好的跟踪效果,而且也有效地推算了囊泡在垂直于成像平面方向上的运动。更多还原

【Abstract】 In this paper, GLUT4 vesicles are observed in real-time under TIRF microscopy and a new three-dimensional single particle tracking algorithm according to the unique features of TIRF is put forward. Firstly a fluorescence correction procedure was processed to solve the problem of fluorescence bleaching over time and mobile vesicles were segmented by an adaptive background subtraction method. Kalman filtering was then introduced to track the granules so as to reduce the searching range and to avoid the disturbance of background noise and false targets. In the experiments the algorithm was applied in analyzing the long-distance movement of GLUT4 vesicles. The experimental results indicate that the algorithm has achieved robust tracking of the vesicles in the imaging plane and has effectively calculated the position in the direction orthogonal to the imaging plane.更多还原