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Ground State Depletion Super-resolution Imaging in Mammalian Cells
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Evaluating the applicability of computational super-resolution techniques for harmonic generation microscopy.

MacAulay Harvey1, Richard Cisek1, Sarry Al-Turk2

  • 1Department of Chemistry, Saint Mary's University, 923 Robie Street, Halifax, NS, B3H 3C3, Canada.

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|June 23, 2026
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Summary

Computational super-resolution techniques enhance imaging resolution for second harmonic generation (SHG) and third harmonic generation (THG) microscopy without specialized equipment. These methods improve nano imaging capabilities and preserve polarization information for advanced SHG microscopy applications.

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Area of Science:

  • Nonlinear Optics
  • Microscopy
  • Nanotechnology

Background:

  • Second harmonic generation (SHG) and third harmonic generation (THG) microscopy are powerful nonlinear optical techniques for imaging nanostructures.
  • Diffraction limits spatial resolution in SHG and THG imaging, hindering nanoscale investigations.
  • Existing super-resolution methods for SHG/THG require nonstandard, costly microscope components.

Purpose of the Study:

  • To investigate the application of computational super-resolution (CSR) techniques for SHG and THG imaging.
  • To enable super-resolution polarization-resolved SHG microscopy using accessible CSR methods.
  • To overcome the technical and financial barriers associated with current super-resolution SHG/THG approaches.

Main Methods:

  • Applied several computational super-resolution algorithms to SHG and THG imaging data.
  • Focused on techniques adaptable to standard laser scanning harmonic generation microscopes.
  • Evaluated the preservation of polarization information in SHG signals using CSR.

Main Results:

  • Achieved a lateral resolution enhancement of up to 3.4× for isolated nanostructures compared to standard SHG/THG microscopy.
  • Demonstrated that CSR improves localization accuracy in harmonic generation microscopy, despite inherent signal ambiguities.
  • Confirmed that specific CSR techniques (super-resolution radial fluctuations, deblurring by pixel reassignment) preserve SHG polarization dependence.

Conclusions:

  • Computational super-resolution offers a viable, cost-effective path to super-resolution SHG and THG imaging.
  • CSR enables super-resolution polarization-resolved SHG microscopy without modifying existing microscope systems.
  • These findings significantly lower the barrier to entry for advanced nano imaging applications using SHG and THG.