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In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse.
Biomed Opt Express
Förster resonance energy transfer (FRET) is a powerful biological tool for reading out cell signaling processes. In vivo use of FRET is challenging because of the scattering properties of bulk tissue. By combining diffuse fluorescence tomography with fluorescence lifetime imaging (FLIM), implemented using wide-field time-gated detection of fluorescence excited by ultrashort laser pulses in a tomographic imaging system and applying inverse scattering algorithms, we can reconstruct the three dimensional spatial localization of fluorescence quantum efficiency and lifetime. We demonstrate in vivo spatial mapping of FRET between genetically expressed fluorescent proteins in live mice read out using FLIM. Following transfection by electroporation, mouse hind leg muscles were imaged in vivo and the emission of free donor (eGFP) in the presence of free acceptor (mCherry) could be clearly distinguished from the fluorescence of the donor when directly linked to the acceptor in a tandem (eGFP-mCherry) FRET construct.
|Title:||In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse.|
|Open access status:||An open access publication|
|Keywords:||(170.2655) Functional monitoring and imaging, (170.3010) Image reconstruction techniques, (170.3650) Lifetime-based sensing, (170.3660) Light propagation in tissues, (170.3880) Medical and biological imaging, (170.6960) Tomography|
|UCL classification:||UCL > School of BEAMS > Faculty of Engineering Science
UCL > School of BEAMS > Faculty of Engineering Science > Computer Science
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