Structured illumination fluorescence microscopy with distorted excitations using a filtered blind-SIM algorithm

Area of Science:

• Microscopy and Imaging Technologies
• Biophysics
• Optical Engineering

Background:

• Structured illumination microscopy (SIM) provides super-resolution fluorescence imaging.
• SIM performance is highly sensitive to excitation pattern aberrations and misalignments.
• Existing methods struggle with distorted illumination patterns.

Purpose of the Study:

• To develop a robust SIM reconstruction algorithm capable of handling strongly distorted illumination patterns.
• To extend the blind-SIM technique for simultaneous sample and illumination reconstruction without prior knowledge.
• To validate the algorithm's performance on both synthetic and experimental data.

Main Methods:

• Implementation of an extended blind-SIM reconstruction algorithm.
• Processing of synthetic and experimental SIM datasets with varying degrees of illumination distortion.
• Comparative analysis of reconstructed images against standard SIM methods.

Main Results:

• The algorithm successfully processed SIM data with significantly distorted illumination patterns.
• Reconstruction quality was comparable to established methods for non-distorted illuminations.
• The developed method produced artifact-free super-resolved images even under severe optical aberrations.

Conclusions:

• The novel reconstruction algorithm significantly improves the robustness of SIM imaging.
• This technique enables reliable super-resolution microscopy in the presence of optical imperfections.
• The blind-SIM extension offers a powerful tool for challenging imaging conditions.