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CLC number: TP317.4; Q27

On-line Access: 2008-01-10

Received: 2006-12-29

Revision Accepted: 2007-10-30

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Journal of Zhejiang University SCIENCE A 2008 Vol.9 No.2 P.232~240


Three-dimensional tracking of GLUT4 vesicles in TIRF microscopy

Author(s):  Xiang-ping WU, Jie-yue LI, Ying-ke XU, Ke-di XU, Xiao-xiang ZHENG

Affiliation(s):  Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China; more

Corresponding email(s):   wxp_hzbme@yahoo.com.cn, zxx@mail.bme.zju.edu.cn

Key Words:  GLUT4, Total internal reflection fluorescence (TIRF) microscopy, Adaptive background subtraction, Kalman filter, Fluorescence correction

Xiang-ping WU, Jie-yue LI, Ying-ke XU, Ke-di XU, Xiao-xiang ZHENG. Three-dimensional tracking of GLUT4 vesicles in TIRF microscopy[J]. Journal of Zhejiang University Science A, 2008, 9(2): 232~240.

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author="Xiang-ping WU, Jie-yue LI, Ying-ke XU, Ke-di XU, Xiao-xiang ZHENG",
journal="Journal of Zhejiang University Science A",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Three-dimensional tracking of GLUT4 vesicles in TIRF microscopy
%A Xiang-ping WU
%A Jie-yue LI
%A Ying-ke XU
%A Ke-di XU
%A Xiao-xiang ZHENG
%J Journal of Zhejiang University SCIENCE A
%V 9
%N 2
%P 232~240
%@ 1673-565X
%D 2008
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.A061659

T1 - Three-dimensional tracking of GLUT4 vesicles in TIRF microscopy
A1 - Xiang-ping WU
A1 - Jie-yue LI
A1 - Ying-ke XU
A1 - Ke-di XU
A1 - Xiao-xiang ZHENG
J0 - Journal of Zhejiang University Science A
VL - 9
IS - 2
SP - 232
EP - 240
%@ 1673-565X
Y1 - 2008
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.A061659

TIRF microscopy has provided a means to view mobile granules within 100 nm in size in two dimensions. However quantitative analysis of the position and motion of those granules requires an appropriate tracking method. In this paper, we present a new tracking algorithm combined with the unique features of TIRF. Firstly a fluorescence correction procedure was processed to solve the problem of fluorescence bleaching over time. Mobile granules were then segmented from a time-lapse image stack by an adaptive background subtraction method. kalman filter was introduced to estimate and track the granules that allowed reducing searching range and hence greater reliability in tracking process. After the tracked granules were located in x-y plane, the z-position was indirectly inferred from the changes in their intensities. In the experiments the algorithm was applied in tracking GLUT4 vesicles in living adipose cells. The results indicate that the algorithm has achieved robust estimation and tracking of the vesicles in three dimensions.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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