Wu Xiang-Ping, Li Jie-Yue, Xu Ying-Ke, Xu Ke-Di, Zheng Xiao-Xiang
Dept. of Biomedical Engineering, Key Lab. of Biomedical Engineering, Ministry of Education, Zhejiang University, Hangzhou 310027.
Zhongguo Yi Liao Qi Xie Za Zhi. 2008 Jan;32(1):14-8.
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.
本文在全内反射荧光显微镜（TIRF显微镜）下实时观察葡萄糖转运蛋白4（GLUT4）囊泡，并根据TIRF的独特特征提出了一种新的三维单粒子跟踪算法。首先进行荧光校正程序以解决荧光随时间漂白的问题，通过自适应背景减法方法分割移动囊泡。然后引入卡尔曼滤波来跟踪颗粒，以减小搜索范围并避免背景噪声和虚假目标的干扰。在实验中，该算法被应用于分析GLUT4囊泡的长距离运动。实验结果表明，该算法在成像平面上实现了对囊泡的稳健跟踪，并有效地计算了与成像平面正交方向上的位置。
