Point-spread function engineering to reduce the impact of spherical aberration on 3D computational fluorescence...

Area of Science:

• Optical microscopy
• Computational imaging
• Wavefront engineering

Background:

• Depth-induced spherical aberration (SA) complicates 3D microscopy.
• Computational complexity increases with SA in 3D imaging.
• Existing methods struggle with SA sensitivity.

Purpose of the Study:

• To engineer 3D point-spread functions (PSFs) using wavefront encoding (WFE) to reduce SA sensitivity.
• To evaluate WFE-PSF performance for extended-depth-of-field (EDOF) and computational optical sectioning microscopy (COSM).
• To identify optimal cubic phase mask designs for improved 3D imaging.

Main Methods:

• Investigated WFE with various cubic phase mask designs.
• Engineered 3D PSFs to mitigate SA effects.
• Evaluated PSF sensitivity to defocus and SA using mean-square-error metrics.
• Simulated WFE-microscope images to assess performance.

Main Results:

• Demonstrated SA insensitivity of engineered WFE-PSFs over 30 μm sample depths.
• Identified specific phase mask parameters for reduced SA sensitivity.
• WFE designs showed low sensitivity to SA while maintaining defocus sensitivity for COSM.
• Successful WFE designs were validated through simulated imaging.

Conclusions:

• WFE is effective in engineering SA-insensitive PSFs for 3D microscopy.
• Optimized WFE designs enable robust EDOF and COSM applications.
• This approach reduces computational load and improves imaging quality in challenging conditions.