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

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

Houwink, Quint (Student TU Delft)
Kalisvaart, D. (TU Delft Team Carlas Smith)
Hung, S. (TU Delft Team Carlas Smith)
Cnossen, J.P. (TU Delft Team Carlas Smith)
Fan, D. (TU Delft Team Carlas Smith)
Mos, Paul (EPFL Neuchâtel)
Ülkü, Arin Can (EPFL Neuchâtel)
Bruschini, Claudio (EPFL Neuchâtel)
Charbon-Iwasaki-Charbon, E. (EPFL Neuchâtel)
Smith, C.S. (TU Delft Team Carlas Smith)

2021

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.

http://resolver.tudelft.nl/uuid:977f6003-579a-4ce9-8c79-042941f2b93a

https://doi.org/10.1364/OE.439340

1094-4087

Optics Express, 29 (24), 39920-39929

Institutional Repository

journal article

© 2021 Quint Houwink, D. Kalisvaart, S. Hung, J.P. Cnossen, D. Fan, Paul Mos, Arin Can Ülkü, Claudio Bruschini, E. Charbon-Iwasaki-Charbon, C.S. Smith