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Three-dimensional bi-functional refractive index and fluorescence microscopy (BRIEF).

Yi Xue1, David Ren2, Laura Waller2

  • 1Department of Biomedical Engineering, University of California, Davis, CA 95616, USA.

Biomedical Optics Express
|February 3, 2023
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Summary
This summary is machine-generated.

This study introduces a novel multi-modal microscopy technique to simultaneously capture 3D fluorescence and refractive index (RI) data. This method enables digital correction of scattering effects, improving fluorescence image quality in biological tissues.

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Computational Imaging

Background:

  • Fluorescence microscopy offers molecular specificity but lacks structural information.
  • Phase microscopy provides label-free refractive index (RI) for structural insights.
  • Conventional methods capture either fluorescence or RI, not both simultaneously.

Purpose of the Study:

  • To develop a computational multi-modal imaging method for simultaneous 3D fluorescence and 3D RI reconstruction.
  • To enable digital correction of multiple-scattering effects in fluorescence imaging using RI data.
  • To advance label-free imaging capabilities in biological samples.

Main Methods:

  • Developed a novel computational multi-modal imaging method based on epi-mode microscopy.
  • Acquired multiple fluorescence images illuminated by single fluorophores.
  • Solved an inverse problem using a multiple-scattering forward model to reconstruct 3D RI and fluorescence.

Main Results:

  • Successfully reconstructed both 3D fluorescence and 3D RI from a single dataset.
  • Demonstrated the method's efficacy for epi-mode 3D RI imaging.
  • Showcased digital correction of multiple-scattering effects in fluorescence images.

Conclusions:

  • The developed method enables simultaneous acquisition of functional (fluorescence) and structural (RI) information.
  • This technique can digitally correct scattering-induced image degradation in biological tissues.
  • Offers a powerful new tool for advanced biological imaging and analysis.