title: Recording intracellular cAMP levels with EPAC-based FRET sensors by fluorescence lifetime imaging.
abstract: Eukaryotic cells use second messengers such as Ca(2+), IP3, cGMP, and cAMP to transduce extracellular signals like hormones, via membrane receptors to downstream cellular effectors. FRET-based sensors are ideal to visualize and measure these rapid changes of second messenger concentrations in time and place. Here, we describe the use of EPAC-based FRET sensors to measure cAMP with spatiotemporal resolution in cells by fluorescence lifetime imaging (FLIM).
authors.affiliation: Department of Cell Biology and Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Raspe
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Klarenbeek
authors.forename: Kees
authors.initials: K
authors.lastname: Jalink
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D045744:Cell Line, Tumor; D000242:Cyclic AMP / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D017930:Genes, Reporter; D006801:Humans; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods
chemical_list: D000242:Cyclic AMP