Computational correction of spatially variant optical aberrations in 3D single-molecule localization microscopy

Area of Science:

• Biophysics
• Optical Microscopy
• Computational Imaging

Background:

• 3D single-molecule localization microscopy (SMLM) is crucial for high-resolution biological imaging.
• Accurate localization in SMLM depends on precise point-spread-function (PSF) fitting.
• Optical aberrations distort PSFs, reducing localization accuracy and precision.

Purpose of the Study:

• To develop and validate a computational method for correcting optical aberrations in ultrawide-field SMLM.
• To quantify the impact of spatially varying aberrations on emitter localization.
• To improve the accuracy and precision of 3D emitter localization across the entire field of view.

Main Methods:

• Employing computational phase retrieval based on a vectorial PSF model.
• Quantifying spatial variance of optical aberrations in a two-channel ultrawide-field microscope.
• Applying a spatially variant PSF model for emitter localization.

Main Results:

• Successfully quantified optical aberrations across the ultrawide field of view.
• Demonstrated significant improvement in localization accuracy and precision in x, y, and z directions.
• The vectorial PSF model effectively corrects for aberrations, enhancing SMLM performance.

Conclusions:

• Computational phase retrieval with vectorial PSF modeling is effective for correcting optical aberrations in SMLM.
• This approach enables accurate and precise 3D emitter localization over large imaging areas.
• The method significantly enhances the reliability and applicability of ultrawide-field SMLM for biological research.