Active illumination for wide-field time-resolved fluorescence imaging

Lingling Zhao; Ken Abe; Margarida Barroso; Xavier Intes

doi:10.1117/12.2033425

18 June 2013 Active illumination for wide-field time-resolved fluorescence imaging

Lingling Zhao, Ken Abe, Margarida Barroso, Xavier Intes

Proceedings Volume 8801, Novel Biophotonic Techniques and Applications II; 88010D (2013) https://doi.org/10.1117/12.2033425
Event: European Conferences on Biomedical Optics, 2013, Munich, Germany

Abstract

The large dynamic range of fluorescence emission collected is one of the major challenges in wide-field fluorescence lifetime imaging. To overcome this challenge, we developed an active illumination strategy to acquire optimal fluorescence signals over the sample imaged even in the presence of large fluorophore concentration distributions. We validated the stability of our approach in a multi-well plate setting with fluorophore concentrations ranging <2 orders of magnitude. We report the ability of our method to retrieve accurately the lifetime over this concentration range based on optimized wide-field data. Our results demonstrate that active wide-field illumination can improve the signal-to-noise ratio and weak-signal sensitivity for enhanced accuracy of fluorescence decay curve fitting and lifetime estimation at high acquisition speed.

Citation Download Citation

Lingling Zhao, Ken Abe, Margarida Barroso, and Xavier Intes "Active illumination for wide-field time-resolved fluorescence imaging", Proc. SPIE 8801, Novel Biophotonic Techniques and Applications II, 88010D (18 June 2013); https://doi.org/10.1117/12.2033425

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