TIRF显微镜下GLUT4囊泡的三维运动分析  

Three-Dimensional Motion Analysis for GLUT4 Vesicles in TIRF Microscopy

作　　者：吴向平[1] 李届悦[1] 许迎科[1] 许科帝[1] 郑筱祥[1] 

机构地区：[1]浙江大学生物医学工程系生物医学工程教育部重点实验室,浙江杭州310027

出　　处：《中国医疗器械杂志》2008年第1期14-18,共5页Chinese Journal of Medical Instrumentation

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。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.

关 键 词：TIRF显微镜 GLUT4 单粒子跟踪 荧光校正 KALMAN滤波 

分 类 号：R318[医药卫生—生物医学工程]