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Theoretical minimum uncertainty of single-molecule localizations using a single-photon avalanche diode array.

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    Summary
    This summary is machine-generated.

    We derived a binomial Cramér-Rao lower bound (CRLB) for single-photon avalanche diode (SPAD) arrays used in single-molecule localization microscopy (SMLM). This new CRLB accurately predicts localization uncertainty for SPAD-based SMLM across various emitter intensities.

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    Area of Science:

    • Biophysics
    • Optical Microscopy
    • Photonics

    Background:

    • Single-photon avalanche diode (SPAD) arrays offer high frame rates and zero readout noise, making them suitable for single-molecule localization microscopy (SMLM).
    • SPAD arrays produce binary frame outputs, necessitating a departure from the standard Poissonian model for photon detection.

    Purpose of the Study:

    • To derive and validate a binomial Cramér-Rao lower bound (CRLB) for localization uncertainty in SMLM using SPAD arrays.
    • To address the limitations of the Poissonian CRLB in predicting localization precision with SPAD detectors.

    Main Methods:

    • Derivation of a theoretical binomial CRLB applicable to SPAD detector characteristics.
    • Benchmarking the binomial CRLB against simulated and experimental SMLM data.
    • Comparison of localization uncertainty predictions between binomial and Poissonian CRLB models.

    Main Results:

    • The derived binomial CRLB accurately models localization uncertainty for SPAD-based SMLM.
    • Under specific conditions (photon count > 1 within PSF std dev), the binomial CRLB indicates a 46% higher theoretical uncertainty than the Poissonian CRLB.
    • For typical SMLM photon fluxes without saturation, the binomial CRLB aligns with the Poissonian CRLB predictions.

    Conclusions:

    • The binomial CRLB is essential for accurate prediction and benchmarking of localization uncertainty in SMLM utilizing SPAD arrays.
    • This framework is applicable for all practical emitter intensities in SMLM experiments with SPADs.