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Optics Letters

Optics Letters


  • Editor: Alan E. Willner
  • Vol. 38, Iss. 19 — Oct. 1, 2013
  • pp: 3976–3979

Active wide-field illumination for high-throughput fluorescence lifetime imaging

Lingling Zhao, Ken Abe, Margarida Barroso, and Xavier Intes  »View Author Affiliations

Optics Letters, Vol. 38, Issue 19, pp. 3976-3979 (2013)

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Wide-field fluorescence lifetime imaging allows for fast imaging of large sample areas at the cost of low sensitivity to weak fluorescence signals. To overcome this challenge, we developed an active wide-field illumination (AWFI) strategy to optimize the impinging spatial intensity for acquiring optimal fluorescence signals over the whole sample. We demonstrated the ability of AWFI to accurately estimate lifetimes from a multiwell plate sample with concentrations ranging over two orders of magnitude. We further reported its successful application to a quantitative Förster resonance energy transfer lifetime cell-based assay. Overall, this method allows for enhanced accuracy in lifetime-based imaging at high acquisition speed over samples with large fluorescence intensity distributions.

© 2013 Optical Society of America

OCIS Codes
(110.3080) Imaging systems : Infrared imaging
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(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
(170.2945) Medical optics and biotechnology : Illumination design

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: April 9, 2013
Revised Manuscript: August 19, 2013
Manuscript Accepted: September 9, 2013
Published: September 30, 2013

Lingling Zhao, Ken Abe, Margarida Barroso, and Xavier Intes, "Active wide-field illumination for high-throughput fluorescence lifetime imaging," Opt. Lett. 38, 3976-3979 (2013)

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