title: Imaging changes in cytoplasmic calcium using the Yellow Cameleon 3.6 biosensor and confocal microscopy.
abstract: Changes in the concentration of cytoplasmic calcium, [Ca(2+)]cyt are central regulators in many cellular signal transduction pathways including many lipid-mediated regulatory networks. Given this central role that [Ca(2+)] has during plant growth, monitoring spatial and temporal [Ca(2+)] dynamics can reveal a critical component of cellular physiology. Here, we describe the measurement of [Ca(2+)]cyt in Arabidopsis root cells using plants expressing Yellow Cameleon 3.6 (YC 3.6). YC3.6 is a Ca(2+)-sensitive biosensor where the intensity of its fluorescence resonance energy transfer (FRET) signal changes as the Ca(2+) 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 Ca(2+) using an approach called ratio analysis.
authors.affiliation: Department of Botany, University of Wisconsin, Madison, WI, USA.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Swanson
authors.forename: Simon
authors.initials: S
authors.lastname: Gilroy
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002138:Calibration; D003593:Cytoplasm / Q000378:metabolism*; D018613:Microscopy, Confocal / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D018517:Plant Roots / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; C495639:yellow cameleon; D002118:Calcium