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Real-time image resolution measurement for single molecule localization microscopy.

Mengting Li, Mingtao Shang, Luchang Li

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    This study introduces a real-time method to measure image resolution in single molecule localization microscopy (SMLM). This technique optimizes imaging throughput while ensuring consistent resolution across heterogeneous samples.

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    Area of Science:

    • Microscopy
    • Biotechnology
    • Image Analysis

    Background:

    • Single molecule localization microscopy (SMLM) shows promise for high-throughput screening.
    • Optimizing imaging throughput without compromising image quality, particularly resolution homogeneity, remains a challenge for SMLM in heterogeneous samples.

    Purpose of the Study:

    • To develop a real-time image resolution measurement method for SMLM.
    • To address the limitation of maintaining homogeneous resolution during high-throughput imaging of multiple fields of view (FOVs).

    Main Methods:

    • Introduced a real-time image resolution measurement method for SMLM.
    • Employed a heuristic framework for overall image resolution, considering localization precision and density.
    • Utilized the spatial Poisson process to model molecule activation and determine real-time localization density, avoiding post-acquisition estimation.

    Main Results:

    • Validated the method for real-time resolution measurement in SMLM.
    • Demonstrated effective guarantee of homogeneous image resolution across multiple FOVs.
    • Achieved optimized imaging throughput without sacrificing image quality.

    Conclusions:

    • The developed real-time method enables efficient and high-quality SMLM imaging.
    • This approach is effective for heterogeneous samples, ensuring consistent resolution and optimized throughput.
    • Facilitates broader applications of SMLM in high-throughput screening.