Quenched Flow Analysis of Exocytosis in Paramecium Cells: Time Course, Changes in Membrane Structure, and Calcium Requirements Revealed after Rapid Mixing and Rapid Freezing of Intact Cells

Synchronous exocytosis in Paramecium cells was analyzed on a subsecond time scale. For this purpose we developed a quenched flow device for rapid mixing and rapid freezing of cells without impairment (time resolution in the millisecond range, dead time ∼30 ms). Cells frozen at defined times after st...

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Bibliographic Details
Published in:The Journal of cell biology 1991-06, Vol.113 (6), p.1295-1304
Main Authors: Knoll, Gerd, Braun, Claudia, Plattner, Helmut
Format: Article
Language:English
Subjects:
Acinar cells
Animals
Biological and medical sciences
Calcium
Calcium - metabolism
Cell Membrane - metabolism
Cell Membrane - ultrastructure
Cell membranes
Cell physiology
Cells
Chromaffin cells
Defibrillators
Exocytosis
Freeze Fracturing - methods
freeze-fracture
Freezing
Fundamental and applied biological sciences. Psychology
Kinetics
Mast cells
Membrane Fusion
membrane structure
Microscopy, Electron
Molecular and cellular biology
P branes
Paramecium - metabolism
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Summary:Synchronous exocytosis in Paramecium cells was analyzed on a subsecond time scale. For this purpose we developed a quenched flow device for rapid mixing and rapid freezing of cells without impairment (time resolution in the millisecond range, dead time ∼30 ms). Cells frozen at defined times after stimulation with the noncytotoxic secretagogue aminoethyldextran were processed by freeze substitution for electron microscopic analysis. With ultrathin sections the time required for complete extrusion of secretory contents was determined to be
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.113.6.1295