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Compressive hyperspectral time-resolved wide-field fluorescence lifetime imaging


Spectrally resolved fluorescence lifetime imaging1,2,3 and spatial multiplexing1,4,5 have offered information content and collection-efficiency boosts in microscopy, but efficient implementations for macroscopic applications are still lacking. An imaging platform based on time-resolved structured light and hyperspectral single-pixel detection has been developed to perform quantitative macroscopic fluorescence lifetime imaging (MFLI) over a large field of view (FOV) and multiple spectral bands simultaneously. The system makes use of three digital micromirror device (DMD)-based spatial light modulators (SLMs) to generate spatial optical bases and reconstruct N by N images over 16 spectral channels with a time-resolved capability (40 ps temporal resolution) using fewer than N2 optical measurements. We demonstrate the potential of this new imaging platform by quantitatively imaging near-infrared (NIR) Förster resonance energy transfer (FRET) both in vitro and in vivo. The technique is well suited for quantitative hyperspectral lifetime imaging with a high sensitivity and paves the way for many important biomedical applications.

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Figure 1: System diagram and typical hyperspectral TPSF example.
Figure 2: Results of in vitro study.
Figure 3: Results of the in vivo study.


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This work is supported by the National Science Foundation through Career Award CBET 1149407, and National Institute of Health Grants R01 EB19443 and R01 CA207725. The authors acknowledge the support from M. Barroso and A. Rudkouskaya for the sample preparation in animal studies, and the insightful discussions with W. Cong and G. Wang.

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Authors and Affiliations



X.I. conceived the original idea. Q.P. contributed to the platform instrumentation, control software design and experimental validation. R.Y. contributed to the pattern implementation, initial simulations and image reconstruction. N.S. contributed to the animal study protocol and experiments. All the authors contributed to the analysis of the experimental results and to writing the manuscript.

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Correspondence to Xavier Intes.

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The authors declare no competing financial interests.

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Pian, Q., Yao, R., Sinsuebphon, N. et al. Compressive hyperspectral time-resolved wide-field fluorescence lifetime imaging. Nature Photon 11, 411–414 (2017).

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