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Measuring Ca²⁺ sparks in cardiac myocytes.

Authors: Macquaide, Niall; Bito, Virginie; Sipido, Karin R.;

Measuring Ca²⁺ sparks in cardiac myocytes.

Abstract

This protocol describes the measurement of Ca(2+) sparks in intact myocytes by using a Ca(2+)-sensitive dye and imaging using laser scanning confocal microscopy. It takes advantage of spontaneous Ca(2+)-release events-sparks-using them as a measure of the activity of ryanodine receptors (RyRs). Two methodologies are described: One requires that cardiomyocytes be stimulated, preferably under voltage clamp by depolarizing pulses, until steady-state is reached, and then stimulation is stopped and Ca(2+) sparks are recorded. The second requires that cells be permeabilized and bathed in a solution to load the cell with Ca(2+) sufficient to elicit Ca(2+) sparks, but not Ca(2+) waves. These are then analyzed offline to quantify spark frequency and morphology. The advantages and disadvantages of each approach are discussed.

Country
United Kingdom
Related Organizations
Keywords

Cytological Techniques/methods, Microscopy, Myocytes, Microscopy, Confocal, Patch-Clamp Techniques, Calcium/analysis, Staining and Labeling, Patch-Clamp Techniques/methods, Cytological Techniques, Cardiac/chemistry, Genetics and Molecular Biology, Confocal/methods, Fluorescent Dyes/metabolism, General Biochemistry, Calcium, Myocytes, Cardiac, Staining and Labeling/methods, Fluorescent Dyes

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Powered by OpenAIRE graph
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
2
Average
Average
Average
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