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Measurement precision bounds on aberrated single-molecule emission patterns.

Li Fang, Fang Huang

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    |November 22, 2024
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    Summary

    Optical aberrations limit precision in single-molecule localization microscopy (SMLM). This study quantifies aberration effects on 2D and 3D SMLM, providing guidelines for aberration correction to improve imaging reliability.

    Area of Science:

    • Optics
    • Biophysics
    • Microscopy

    Background:

    • Single-molecule localization microscopy (SMLM) offers nanoscale resolution but is degraded by optical aberrations.
    • These aberrations distort point spread functions (PSFs), reducing localization precision and 3D imaging accuracy.
    • A comprehensive analysis of various aberration types' impact on SMLM is lacking.

    Purpose of the Study:

    • To quantitatively analyze the theoretical precision limits for position and wavefront measurements under optical aberrations in SMLM.
    • To compare the effects of different aberration types on localization precision in 2D, biplane, and astigmatism 3D SMLM modalities.
    • To assess wavefront estimation precision from aberrated single-molecule patterns for adaptive optics.

    Main Methods:

    • Utilized Fisher information and Cramér-Rao lower bound (CRLB) for theoretical precision analysis.

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  • Simulated and analyzed the impact of various aberrations, including index mismatch, on SMLM.
  • Evaluated localization precision in different 3D SMLM techniques and 2D imaging.
  • Investigated wavefront estimation accuracy from aberrated PSFs.

    • Main Results:

    • Quantified the theoretical limits of localization and wavefront sensing precision in the presence of aberrations.
    • Demonstrated how different aberration types differentially impact localization precision across 2D and 3D SMLM methods.
    • Showcased the feasibility of precise wavefront estimation for adaptive optics applications in SMLM.
    • Identified specific aberration effects crucial for improving SMLM performance.

    Conclusions:

    • Optical aberrations significantly affect SMLM precision, with varying impacts depending on aberration type and imaging modality.
    • Quantitative analysis provides a foundation for developing effective aberration correction strategies.
    • This work guides the advancement of SMLM for imaging complex biological systems like whole cells and tissues.
    • Enhanced aberration correction will improve the reliability and precision of 3D SMLM.