title: Multi-channel imaging of cellular signaling: interplay of Ca2+ and conventional protein kinase C.
abstract: To investigate the coupling between the calcium (Ca(2+)) signal and the conventional protein kinase C alpha (PKCα) translocation, which are both transient processes, a sectioning imaging technique with sufficient scanning speed is required. Here, we describe how to use a Nipkow-disk-based confocal system with an image splitter for two cameras to acquire simultaneously both the Ca(2+) signal and images of PKCα fused to the fluorescent protein DsRed2 in HEK293 cells on ATP stimulation.
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
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: Xin
authors.initials: X
authors.lastname: Hui
authors.affiliation: Cell Biology and Biophysics Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Gergor
authors.initials: G
authors.lastname: Reither
authors.affiliation: Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, School of Medicine, Saarland University, 66421 Homburg/Saar, Germany; Cell Biology and Biophysics Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Kaestner
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D020013:Calcium Signaling; D057809:HEK293 Cells; D006801:Humans; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods*; D061848:Optical Imaging / Q000295:instrumentation* / Q000379:methods*; D051571:Protein Kinase C-alpha / Q000378:metabolism*; D021381:Protein Transport
chemical_list: D000255:Adenosine Triphosphate; D051571:Protein Kinase C-alpha