Abstract
Changes in the concentration of cytoplasmic calcium, [Ca2+]cyt are central regulators in many cellular signal transduction pathways including many lipid-mediated regulatory networks. Given this central role that [Ca2+] has during plant growth, monitoring spatial and temporal [Ca2+] dynamics can reveal a critical component of cellular physiology. Here, we describe the measurement of [Ca2+]cyt in Arabidopsis root cells using plants expressing Yellow Cameleon 3.6 (YC 3.6). YC3.6 is a Ca2+-sensitive biosensor where the intensity of its fluorescence resonance energy transfer (FRET) signal changes as the Ca2+ level within the cell rises and falls. The FRET from this calcium reporter can be visualized using confocal microscopy and the resultant images converted to a quantitative map of the levels of Ca2+ using an approach called ratio analysis.
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Acknowledgments
This work was supported by grants from the USDA (2007-35304-18327), NSF (MCB 0641288) and NASA (NNX09AK80G) to S.G.
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Swanson, S.J., Gilroy, S. (2013). Imaging Changes in Cytoplasmic Calcium Using the Yellow Cameleon 3.6 Biosensor and Confocal Microscopy. In: Munnik, T., Heilmann, I. (eds) Plant Lipid Signaling Protocols. Methods in Molecular Biology, vol 1009. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-401-2_27
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DOI: https://doi.org/10.1007/978-1-62703-401-2_27
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