Photon count estimation in single-molecule localization microscopy

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Photon count estimation in single-molecule localization microscopy

Preprint (2018)

Author(s)

Rasmus Thorsen (TU Delft - ImPhys/Quantitative Imaging)

Christiaan Hulleman (TU Delft - ImPhys/Quantitative Imaging)

Hammer Mathias (University of Massachusetts Medical School)

David Grunwald (University of Massachusetts Medical School)

Sjoerd Stallinga (TU Delft - Applied Sciences)

Bernd Rieger (TU Delft - ImPhys/Quantitative Imaging)

Research Group

ImPhys/Quantitative Imaging

DOI related publication

https://doi.org/10.1101/396424 Final published version

To reference this document use

https://resolver.tudelft.nl/uuid:16f8cad4-6674-4679-82fc-b44bc490bf2b

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Publication Year

2018

Language

English

Research Group

ImPhys/Quantitative Imaging

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Abstract

Recently, Franke, Sauer and van de Linde introduced a way to estimate the axial position of single-molecules (TRABI). To this end, they compared the detected photon count from a temporal radial-aperture-based intensity estimation to the estimated count from Gaussian point-spread function (PSF) fitting to the data. Empirically they found this photometric ratio to be around 0.7-0.8 close to focus and decreasing away from it. Here, we explain this reported but unexplained discrepancy and furthermore show that the photometric ratio as indicator for axial position is susceptible even to typical optical aberrations.