4.3 Article

Multi-dimensional resolution of elementary Ca2+ signals by simultaneous multi-focal imaging

期刊

CELL CALCIUM
卷 43, 期 4, 页码 367-374

出版社

ELSEVIER SCI LTD
DOI: 10.1016/j.ceca.2007.07.002

关键词

Ca2+ puffs; multi-focal imaging; Ca2+ imaging

资金

  1. 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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