title: Confocal FLIM of genetically encoded FRET sensors for quantitative Ca2+ imaging.
abstract: Fluorescence lifetime imaging (FLIM) is a powerful imaging mode that can be combined with confocal imaging. Changes in the fluorescence decay time of a donor in an intramolecular Förster resonance energy transfer (FRET)-based biosensor provide intrinsic quantitative data. Here, we describe a protocol using both the Ca(2+) sensor TN-XL, which uses troponin C, as the Ca(2+)-sensing unit, and the FLIM technology based on time-correlated single-photon counting.
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: Benjamin
authors.initials: B
authors.lastname: Sauer
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: 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: Lars
authors.initials: L
authors.lastname: Kaestner
mesh_terms: D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D017785:Photons; D014162:Transfection
chemical_list: D002118:Calcium