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Intravital FRET: Probing Cellular and Tissue Function in Vivo |
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| Authors: | Helena Radbruch Daniel Bremer Ronja Mothes Robert Günther Jan Leo Rinnenthal Julian Pohlan Carolin Ulbricht Anja E. Hauser Raluca Niesner |
| Affiliation: | 1.Neuropathology, Charité–University of Medicine, Berlin 10117, Germany; E-Mails: (H.R.); (R.M.); (J.L.R.); (J.P.);2.Germany German Rheumatism Research Center, Berlin 10117, Germany; E-Mails: (D.B.); (R.G.); (C.U.); (A.E.H.);3.Immundynamics and Intravital Microscopy, Charité–University of Medicine, Berlin 10117, Germany |
| 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. |
| Keywords: | intravital FRET multi-photon microscopy fluorescence lifetime imaging genetically encoded calcium indicators |