Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10926593 | Cell Calcium | 2005 | 17 Pages |
Abstract
Recent developments in microscopy and fluorescent indicators now make it possible to monitor the activity and localization of membrane ion channels by imaging Ca2+ flux through individual channels. Such optical approaches have advantages over electrophysiological single-channel techniques in that they are less invasive, provide spatial information and can simultaneously and independently monitor hundreds of channels. However, their kinetic resolution does not yet approach that of patch-clamp recordings. To help understand the processes that determine the temporal resolution and noise level of single-channel Ca2+ fluorescence signals (SCCaFTs), we simulated the microdomains of Ca2+ ions and Ca2+-bound indicator dye that exist around the mouth of an open channel. Further, as an aid to development of improved optical techniques, we modeled the dependence of the amplitude and kinetics of SCCaFTs on parameters such as the imaging volume, the indicator concentration, affinity and mobility, and the presence of endogenous and exogenous Ca2+ buffers. The results indicate that under optimal conditions, including the use of confocal or total-internal reflection microscopy to image from sub-femtolitre volumes, SCCaFTs should resolve channel openings as brief as 1Â ms with a signal-to-noise ratio >10.
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Authors
Jianwei Shuai, Ian Parker,