title: Intravital FRET: Probing Cellular and Tissue Function in Vivo.
abstract: The development of intravital Förster Resonance Energy Transfer (FRET) is required to probe cellular and tissue function in the natural context: the living organism. Only in this way can biomedicine truly comprehend pathogenesis and develop effective therapeutic strategies. Here we demonstrate and discuss the advantages and pitfalls of two strategies to quantify FRET in vivo-ratiometrically and time-resolved by fluorescence lifetime imaging-and show their concrete application in the context of neuroinflammation in adult mice.
authors.affiliation: Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. Helena.radbruch@charite.de.
authors.forename: Helena
authors.initials: H
authors.lastname: Radbruch
authors.affiliation: Germany German Rheumatism Research Center, Berlin 10117, Germany. daniel.bremer@drfz.de.
authors.forename: Daniel
authors.initials: D
authors.lastname: Bremer
authors.affiliation: Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. ronja.mothes@charite.de.|Germany German Rheumatism Research Center, Berlin 10117, Germany. ronja.mothes@charite.de.
authors.forename: Ronja
authors.initials: R
authors.lastname: Mothes
authors.affiliation: Germany German Rheumatism Research Center, Berlin 10117, Germany. r.guenther@drfz.de.
authors.forename: Robert
authors.initials: R
authors.lastname: Günther
authors.affiliation: Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. jan-leo.rinnenthal@charite.de.
authors.forename: Jan Leo
authors.initials: JL
authors.lastname: Rinnenthal
authors.affiliation: Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. Julian.pohlan@charite.de.|Germany German Rheumatism Research Center, Berlin 10117, Germany. Julian.pohlan@charite.de.
authors.forename: Julian
authors.initials: J
authors.lastname: Pohlan
authors.affiliation: Germany German Rheumatism Research Center, Berlin 10117, Germany. carolin.ulbricht@drfz.de.|Immundynamics and Intravital Microscopy, Charité-University of Medicine, Berlin 10117, Germany. carolin.ulbricht@drfz.de.
authors.forename: Carolin
authors.initials: C
authors.lastname: Ulbricht
authors.affiliation: Germany German Rheumatism Research Center, Berlin 10117, Germany. hauser@drfz.de.|Immundynamics and Intravital Microscopy, Charité-University of Medicine, Berlin 10117, Germany. hauser@drfz.de.
authors.forename: Anja E
authors.initials: AE
authors.lastname: Hauser
authors.affiliation: Germany German Rheumatism Research Center, Berlin 10117, Germany. niesner@drfz.de.
authors.forename: Raluca
authors.initials: R
authors.lastname: Niesner
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000473:pathology*; D002118:Calcium / Q000032:analysis*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000473:pathology*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D000069416:Intravital Microscopy / Q000379:methods*; D051379:Mice; D061848:Optical Imaging / Q000379:methods*
chemical_list: D002118:Calcium
keywords: fluorescence lifetime imaging; genetically encoded calcium indicators; intravital FRET; multi-photon microscopy