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Int J Mol Sci. 2015 May 21;16(5):11713-27. doi: 10.3390/ijms160511713.

Intravital FRET: Probing Cellular and Tissue Function in Vivo.

Author information

1
Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. Helena.radbruch@charite.de.
2
Germany German Rheumatism Research Center, Berlin 10117, Germany. daniel.bremer@drfz.de.
3
Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. ronja.mothes@charite.de.
4
Germany German Rheumatism Research Center, Berlin 10117, Germany. ronja.mothes@charite.de.
5
Germany German Rheumatism Research Center, Berlin 10117, Germany. r.guenther@drfz.de.
6
Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. jan-leo.rinnenthal@charite.de.
7
Neuropathology, Charité-University of Medicine, Berlin 10117, Germany. Julian.pohlan@charite.de.
8
Germany German Rheumatism Research Center, Berlin 10117, Germany. Julian.pohlan@charite.de.
9
Germany German Rheumatism Research Center, Berlin 10117, Germany. carolin.ulbricht@drfz.de.
10
Immundynamics and Intravital Microscopy, Charité-University of Medicine, Berlin 10117, Germany. carolin.ulbricht@drfz.de.
11
Germany German Rheumatism Research Center, Berlin 10117, Germany. hauser@drfz.de.
12
Immundynamics and Intravital Microscopy, Charité-University of Medicine, Berlin 10117, Germany. hauser@drfz.de.
13
Germany German Rheumatism Research Center, Berlin 10117, Germany. niesner@drfz.de.

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:

fluorescence lifetime imaging; genetically encoded calcium indicators; intravital FRET; multi-photon microscopy

PMID:
26006244
PMCID:
PMC4463726
DOI:
10.3390/ijms160511713
[Indexed for MEDLINE]
Free PMC Article

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