Open AccessTheoretical minimum uncertainty of single-molecule localizations using a single-photon avalanche diode array
Author(s) -
Quint Houwink,
Dylan Kalisvaart,
Shih-Te Hung,
Jelmer Cnossen,
Daniel Fan,
Paul Mos,
Arin Can Ülkü,
Claudio Bruschini,
Edoardo Charbon,
Carlas Smith
Publication year - 2021
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.439340
Subject(s) - cramér–rao bound , physics , optics , photon , photon counting , upper and lower bounds , gaussian , computational physics , mathematics , mathematical analysis , quantum mechanics
Single-photon avalanche diode (SPAD) arrays can be used for single-molecule localization microscopy (SMLM) because of their high frame rate and lack of readout noise. SPAD arrays have a binary frame output, which means photon arrivals should be described as a binomial process rather than a Poissonian process. Consequentially, the theoretical minimum uncertainty of the localizations is not accurately predicted by the Poissonian Cramér-Rao lower bound (CRLB). Here, we derive a binomial CRLB and benchmark it using simulated and experimental data. We show that if the expected photon count is larger than one for all pixels within one standard deviation of a Gaussian point spread function, the binomial CRLB gives a 46% higher theoretical uncertainty than the Poissonian CRLB. For typical SMLM photon fluxes, where no saturation occurs, the binomial CRLB predicts the same uncertainty as the Poissonian CRLB. Therefore, the binomial CRLB can be used to predict and benchmark localization uncertainty for SMLM with SPAD arrays for all practical emitter intensities.