Journal
CELL CALCIUM
Volume 43, Issue 4, Pages 367-374Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.ceca.2007.07.002
Keywords
Ca2+ puffs; multi-focal imaging; Ca2+ imaging
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Funding
- NIGMS NIH HHS [R01 GM048071, GM 48071, R01 GM048071-17, R37 GM048071] Funding Source: Medline
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Elementary events such as puffs and sparks are cytosolic microdomains of Ca2+ from which cellular Ca2+ signals are constructed. Because of the tight localization and fast kinetics of elementary events, imaging studies have been hindered by instrumental limitations of confocal and deconvolution fluorescence microscopy which necessitate compromises between spatial and temporal resolution. Here, we describe a novel, yet simple 'multi-focal' fluorescence microscopy system that employs three high-speed cameras focused at different axial depths to enable 4-dimensional imaging with millisecond resolution. We demonstrate the utility of this system for studies of puffs in Xenopus oocytes by mapping the axial distribution of puff sites, by obtaining measurements of puff amplitudes undistorted by focus error, and by deriving deblurred images that reveal novel sub-micron jumps of Ca2+ release sites. Published by Elsevier Ltd.
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