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Tracking of secretory vesicles of PC12 cells by total internal reflection fluorescence microscopy
Author(s) -
Yang D.M.,
Huang C.C.,
Lin H.Y.,
Tsai D.P.,
Kao L.S.,
Chi C.W.,
Lin C.C.
Publication year - 2003
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.2003.01129.x
Subject(s) - total internal reflection fluorescence microscope , vesicle , fluorescence microscope , secretory vesicle , exocytosis , microscopy , biophysics , fluorescence , vesicle fusion , microbiology and biotechnology , chemistry , membrane , biology , biochemistry , optics , synaptic vesicle , physics
Summary Total internal reflection fluorescence microscopy is used to detect cellular events near the plasma membrane. Behaviours of secretory vesicles near the cell surface of living PC12 cells, a neuroendocrine cell line, are studied. The secretory vesicles are labelled by over‐expression of enhanced green fluorescent protein‐tagged Rab3A, one of the small G proteins involved in the fusion of secretory vesicles to plasma membrane in PC12 cells. Images acquired by a fast cooled charge‐coupled device camera using conventional fluorescence microscopy and total internal reflection fluorescence microscopy are compared and analysed. Within the small evanescent range (< 200 nm), the movements of the secretory vesicles of PC12 cells before and after stimulation by high K + are examined. The movements of one vesicle relative to another already docked on the membrane are detected. Total internal reflection fluorescence microscopy provides a novel optical method to trace and analyse the exocytotic events and vesicle specifically near a cell membrane without interference of signals from other parts of the cell.