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Compressive hyperspectral time-resolved wide-field fluorescence lifetime imaging.

Qi Pian1, Ruoyang Yao1, Nattawut Sinsuebphon1

  • 1Biomedical Engineering Department, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York 12180, USA.

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Summary

A new macroscopic fluorescence lifetime imaging (MFLI) system uses time-resolved structured light and hyperspectral detection. This enables efficient, quantitative imaging over large areas, advancing biomedical applications.

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Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Spectroscopy

Background:

  • Fluorescence lifetime imaging (FLIM) and spatial multiplexing enhance microscopy but lack efficient macroscopic implementations.
  • Current techniques struggle with large fields of view and simultaneous multi-spectral data acquisition.

Purpose of the Study:

  • To develop an efficient platform for quantitative macroscopic fluorescence lifetime imaging (MFLI) over a large field of view (FOV) and multiple spectral bands.
  • To enable time-resolved hyperspectral imaging with high sensitivity for biomedical applications.

Main Methods:

  • Developed a system using time-resolved structured light and hyperspectral single-pixel detection.
  • Employed three digital micromirror device (DMD)-based spatial light modulators (SLMs) for spatial optical basis generation.
  • Achieved reconstruction of N x N images over 16 spectral channels with ~40 ps temporal resolution using < N^2 measurements.

Main Results:

  • Successfully demonstrated quantitative macroscopic fluorescence lifetime imaging (MFLI).
  • Successfully imaged near-infrared (NIR) Förster resonance energy transfer (FRET) both in vitro and in vivo.
  • Achieved high sensitivity in quantitative hyperspectral lifetime imaging.

Conclusions:

  • The developed MFLI platform offers efficient, quantitative, and hyperspectral imaging over large FOVs.
  • The technique shows significant potential for sensitive biomedical applications, including FRET imaging.
  • This advancement paves the way for novel diagnostic and research tools in medicine.