Application Notes

Distortion Correction Technique with Non-linear Deconvolution Using Fluorescent Beads as Reference Data

July 2025

Image Scanning Microscopy (ISM) improves spatial resolution without compromising signal intensity. When combined with deconvolution processing, ISM enables the generation of super-resolution images that surpass the traditional optical resolution limits. However, in three-dimensional (3D) imaging, refractive index mismatches between the immersion medium of the objective lens (e.g., water, silicone, oil) and the biological sample can introduce distortions along the optical axis. This remains a challenge even with ISM enhanced in-plane resolution.

In this Application Note, we applied Non-linear Deconvolution using point spread functions (PSFs) extracted from fluorescent beads embedded at various depths. We compared the resulting optical axial resolution with that obtained through standard Linear Deconvolution based on theoretical PSF models.