title: Two-dimensional imaging of fast intracellular Ca2+ release.
abstract: Asynchronous release of calcium (Ca(2+))-for example, the generation of Ca(2+) alternans in cardiac myocytes-is a phenomenon important in the development of cardiac arrhythmogenesis. The development of a failure to release Ca(2+) at individual release sites can be regarded as a major contributor to cardiac pathologies such as hypertrophy. Although confocal linescans provide sufficient temporal resolution to investigate the physiological and pathological cardiac excitation-contraction (EC) coupling, linescans can only image ∼1.5% of the cross section of myocytes, which raises doubts about how representative such recordings are, especially in light of nonhomogeneous uncoupling of Ca(2+) channels and ryanodine receptors. Nowadays, the speed of confocal microscopes has been greatly improved, enabling two-dimensional (2D) imaging at sufficient image rates (>100 frames/sec). To understand better the physiological and pathophysiological EC coupling of cardiomyocytes, we describe here a protocol to monitor fast intracellular Ca(2+) signals using fast 2D confocal scanning.
authors.affiliation: Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, School of Medicine, Saarland University, 66421 Homburg/Saar, Germany.
authors.forename: Qinghai
authors.initials: Q
authors.lastname: Tian
authors.affiliation: Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, School of Medicine, Saarland University, 66421 Homburg/Saar, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Kaestner
authors.affiliation: Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, School of Medicine, Saarland University, 66421 Homburg/Saar, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Lipp
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002478:Cells, Cultured; D007091:Image Processing, Computer-Assisted; D042541:Intracellular Space / Q000378:metabolism*; D018613:Microscopy, Confocal; D057054:Molecular Imaging / Q000379:methods*; D032383:Myocytes, Cardiac / Q000378:metabolism; D013997:Time Factors
chemical_list: D002118:Calcium