title: Tracking of secretory vesicles of PC12 cells by total internal reflection fluorescence microscopy.
abstract: 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.
authors.affiliation: Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: D-M
authors.initials: DM
authors.lastname: Yang
authors.forename: C-C
authors.initials: CC
authors.lastname: Huang
authors.forename: H-Y
authors.initials: HY
authors.lastname: Lin
authors.forename: D-P
authors.initials: DP
authors.lastname: Tsai
authors.forename: L-S
authors.initials: LS
authors.lastname: Kao
authors.forename: C-W
authors.initials: CW
authors.lastname: Chi
authors.forename: C-C
authors.initials: CC
authors.lastname: Lin
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005089:Exocytosis; D049452:Green Fluorescent Proteins; D008027:Light; D008164:Luminescent Proteins; D008561:Membrane Fusion; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D016716:PC12 Cells; D051381:Rats; D022142:Secretory Vesicles / Q000737:chemistry* / Q000378:metabolism; D020826:rab3A GTP-Binding Protein / Q000502:physiology
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D020826:rab3A GTP-Binding Protein