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In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse

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Title: In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse
Authors: McGinty, J
Stuckey, DW
Soloviev, VY
Laine, R
Wylezinska-Arridge, M
Wells, DJ
Arridge, SR
French, PMW
Hajnal, JV
Sardini, A
Item Type: Journal Article
Abstract: 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.
Issue Date: 10-Jun-2011
Date of Acceptance: 10-Jun-2011
URI: http://hdl.handle.net/10044/1/70768
DOI: https://dx.doi.org/10.1364/BOE.2.001907
ISSN: 2156-7085
Publisher: Optical Society of America
Start Page: 1907
End Page: 1917
Journal / Book Title: Biomedical Optics Express
Volume: 2
Issue: 7
Copyright Statement: ©2011 Optical Society of America. Users may use, reuse, and build upon the article, or use the article for text or data mining, so long as such uses are for non-commercial purposes and appropriate attribution is maintained. All other rights are reserved.
Keywords: Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemical Research Methods
Optics
Radiology, Nuclear Medicine & Medical Imaging
Biochemistry & Molecular Biology
RESONANCE ENERGY-TRANSFER
CELLS
(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
Notes: McGinty and Stuckey co-first authors
Appears in Collections:Physics
Bioengineering
Photonics
Institute of Clinical Sciences
Faculty of Natural Sciences



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