title: Three-Dimensional Tracking of Single Granules in Living PC-12 Cells Employing TIRFM and WFFM.
abstract: A comparative study was carried out on evaluating the performance of total internal reflection fluorescence microscopy (TIRFM) and deconvolution wide-field fluorescence microscopy (WFFM) in tracking single secretory granules. Both techniques have been applied to follow the three-dimensional mobility of single secretory granules in living neuroendocrine PC-12 cells. Both techniques return the similar result that most acridine orange-labeled granules were found to travel in random and caged diffusion, and only a small fraction of granules traveled in directed diffusion. Furthermore, the size and 3-D diffusion coefficient of secretory granules, obtained by these two imaging techniques, yield the same value. Together, our results demonstrate the potential of the combination TIRFM and WFFM in tracking long-termed motion of granules throughout live whole cells.
authors.affiliation: Institute of Biophysics and Biochemistry, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xiong
authors.forename: Dongdong
authors.initials: D
authors.lastname: Li
authors.forename: Dan
authors.initials: D
authors.lastname: Zhu
authors.forename: Anlian
authors.initials: A
authors.lastname: Qu