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Sub-Diffraction Photoacoustic Microscopy Enabled by a Novel Phase-Shifted Excitation Strategy: A Numerical Study.

George J Tserevelakis1,2

  • 1Department of Biology, University of Crete, 700 13 Heraklion, Crete, Greece.

Sensors (Basel, Switzerland)
|January 28, 2026
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A new phase-shifted dual-beam excitation method for photoacoustic microscopy significantly enhances lateral resolution, achieving sub-diffraction limits for improved biomedical imaging.

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Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Acoustic Physics

Background:

  • Photoacoustic microscopy (PAM) offers label-free imaging but is limited by optical diffraction.
  • Achieving sub-diffraction-limited resolution is crucial for visualizing fine cellular and vascular structures.

Purpose of the Study:

  • To introduce and simulate a novel phase-shifted dual-beam excitation strategy for frequency-domain photoacoustic microscopy (FD-PAM).
  • To demonstrate the capability of this strategy to achieve optical sub-diffraction-limited lateral resolution.
  • To analyze the impact of optical power ratios on excitation confinement and resolution enhancement.

Main Methods:

  • Numerical simulations were performed to model the excitation dynamics of phase-shifted Gaussian and donut beams.
  • The spatial overlapping and phase-shifted intensity modulation of dual beams were analyzed.
  • The full width at half maximum (FWHM) of the excitation region was calculated and compared to conventional methods.

Main Results:

  • The phase-shifted dual-beam excitation strategy reduced the normalized FWHM from 1.177 to 0.828.
  • This resulted in a ~1.42-fold enhancement in lateral resolution compared to Gaussian beam excitation.
  • An optimal optical power ratio of 1.16 was identified for balancing excitation confinement and side-lobe suppression.

Conclusions:

  • The simulated phase-shifted dual-beam excitation provides a viable pathway for experimental FD-PAM systems.
  • This technique holds potential for high-resolution biomedical imaging of subcellular and microvascular structures.
  • It enables the use of cost-effective continuous-wave laser diodes for advanced imaging applications.