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Near-isotropic super-resolution microscopy with axial interference speckle illumination.

Hajun Yoo1, Kwanhwi Ko1, Sukhyeon Ka1

  • 1School of Electrical and Electronic Engineering, Yonsei University, Seoul, Korea.

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|October 20, 2025
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Summary

Axial interference speckle illumination-engineered structured illumination microscopy (AXIS-SIM) enhances axial resolution in super-resolution imaging. This minimal-modification technique achieves isotropic super-resolution without complex optics, improving biological specimen visualization.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • Super-resolution microscopy advances biological imaging but struggles with isotropic resolution across all dimensions.
  • Achieving high axial resolution often requires complex and sensitive optical setups.

Purpose of the Study:

  • Introduce axial interference speckle illumination-engineered structured illumination microscopy (AXIS-SIM) for enhanced axial resolution.
  • Develop a minimal-modification approach for improved 3D super-resolution imaging.

Main Methods:

  • Utilized constructive interference from a back-reflecting mirror for axial resolution enhancement.
  • Implemented AXIS-SIM without additional phase control or complex beam shaping.
  • Achieved lateral and axial resolutions of 108.5 nm and 140.1 nm, respectively.

Main Results:

  • AXIS-SIM demonstrated superior optical sectioning and improved axial resolution beyond conventional 3D-SIM.
  • The method showed robustness against alignment errors and sample-induced aberrations.
  • Enabled high-throughput 3D super-resolution imaging of diverse biological specimens.

Conclusions:

  • AXIS-SIM offers a simplified yet effective approach to achieving isotropic super-resolution.
  • The technique visualizes 3D cell morphology, nanoscale distributions (lysosomes, microtubules), and lysosomal dynamics with enhanced axial clarity.