Title :
Three-Dimensional Tracking of Single Granules in Living PC-12 Cells Employing TIRFM and WFFM
Author :
Jun Xiong ; Dongdong Li ; Dan Zhu ; Anlian Qu
Author_Institution :
Inst. of Biophys. & Biochem., Huazhong Univ. of Sci. & Technol., Wuhan
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
Keywords :
biodiffusion; biological techniques; cellular transport; fluorescence; optical microscopy; TIRFM; WFFM; acridine orange-labeled granules; caged diffusion; deconvolution wide-field fluorescence microscopy; living neuroendocrine PC-12 cells; random diffusion; three-dimensional single granule tracking; total internal reflection fluorescence microscopy; Biomedical optical imaging; Fluorescence; Lighting; Microscopy; Optical refraction; Optical sensors; Optical surface waves; Optical variables control; Pixel; USA Councils; diffusion coefficient; three-dimensional single particle tracking; total internal reflection fluorescence microscopy; wide-field fluorescence microscopy;
Conference_Titel :
Engineering in Medicine and Biology Society, 2005. IEEE-EMBS 2005. 27th Annual International Conference of the
Conference_Location :
Shanghai
Print_ISBN :
0-7803-8741-4
DOI :
10.1109/IEMBS.2005.1616254
