Calcium (Ca2+) is a fundamentally important component of cellular signal transduction. Dynamic changes in the concentration of Ca2+ ([Ca2+]) in the cytoplasm and within organelles are tightly controlled and regulate a diverse array of biological activities, including fertilization, cell division, gene expression, cellular metabolism, protein biosynthesis, secretion, muscle contraction, intercellular communication, and cell death. Measurement of intracellular [Ca2+] is essential to understanding the role of Ca2+ and for defining the underlying regulatory mechanisms in any cellular process. A broad range of synthetic and biosynthetic fluorescent Ca2+ sensors are available that enable the visualization and quantification of subcellular spatio-temporal [Ca2+] gradients. This chapter describes the application of wide-field digitized video fluorescence microfluorometry and confocal microscopy to quantitatively image Ca2+ in cells with high temporal and spatial resolution.
CITATION STYLE
Roe, M. W., Fiekers, J. F., Philipson, L. H., & Bindokas, V. P. (2006). Visualizing calcium signaling in cells by digitized wide-field and confocal fluorescent microscopy. Methods in Molecular Biology (Clifton, N.J.), 319, 37–66. https://doi.org/10.1007/978-1-59259-993-6_3
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