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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 5, Iss. 3 — Mar. 1, 2014
  • pp: 944–960

Spatial light modulator based active wide-field illumination for ex vivo and in vivo quantitative NIR FRET imaging

Lingling Zhao, Ken Abe, Shilpi Rajoria, Qi Pian, Margarida Barroso, and Xavier Intes  »View Author Affiliations

Biomedical Optics Express, Vol. 5, Issue 3, pp. 944-960 (2014)

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Fluorescence lifetime imaging is playing an increasing role in drug development by providing a sensitive method to monitor drug delivery and receptor-ligand interactions. However, the wide dynamic range of fluorescence intensity emitted by ex vivo and in vivo samples presents challenges in retrieving information over the whole subject accurately and quantitatively. To overcome this challenge, we developed an active wide-field illumination (AWFI) strategy based on a spatial light modulator that acquires optimal fluorescence signals by enhancing the dynamic range, signal to noise ratio, and estimation of lifetime-based parameters. We demonstrate the ability of AWFI to estimate Förster resonance energy transfer (FRET) donor fraction from dissected organs with high accuracy (standard deviation <6%) over the whole field of view, in contrast with the homogenous wide-field illumination. We further report its successful application to quantitative FRET imaging in a live mouse. AWFI allows improved detection of weak signals and enhanced quantitative accuracy in ex vivo and in vivo molecular fluorescence quantitative imaging. The technique allows for robust quantitative estimation of the bio-distribution of molecular probes and lifetime-based parameters over an extended imaging field exhibiting a large range of fluorescence intensities and at a high acquisition speed (less than 1 min).

© 2014 Optical Society of America

OCIS Codes
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(170.6920) Medical optics and biotechnology : Time-resolved imaging
(230.6120) Optical devices : Spatial light modulators
(260.2160) Physical optics : Energy transfer
(260.2510) Physical optics : Fluorescence
(170.2945) Medical optics and biotechnology : Illumination design

ToC Category:
Molecular Imaging and Probe Development

Original Manuscript: November 14, 2013
Revised Manuscript: January 20, 2014
Manuscript Accepted: January 31, 2014
Published: February 27, 2014

Lingling Zhao, Ken Abe, Shilpi Rajoria, Qi Pian, Margarida Barroso, and Xavier Intes, "Spatial light modulator based active wide-field illumination for ex vivo and in vivo quantitative NIR FRET imaging," Biomed. Opt. Express 5, 944-960 (2014)

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