Photon count estimation in single-molecule localization microscopy

Rasmus Thorsen; Christiaan Hulleman; Hammer Mathias; David Grunwald; Sjoerd Stallinga; Bernd Rieger

doi:10.1101/396424

Photon count estimation in single-molecule localization microscopy

Rasmus Thorsen, Christiaan Hulleman, Hammer Mathias, David Grunwald, Sjoerd Stallinga, Bernd Rieger

ImPhys/Imaging Physics

Research output: Working paper/Preprint › Preprint

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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.

Original language	English
Number of pages	16
DOIs	https://doi.org/10.1101/396424
Publication status	Published - 2018

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396424.fullSubmitted manuscript, 2.26 MBLicence: CC BY-NC

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title = "Photon count estimation in single-molecule localization microscopy",

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.",

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T1 - Photon count estimation in single-molecule localization microscopy

AU - Thorsen, Rasmus

AU - Hulleman, Christiaan

AU - Mathias, Hammer

AU - Grunwald, David

AU - Stallinga, Sjoerd

AU - Rieger, Bernd

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

U2 - 10.1101/396424

DO - 10.1101/396424

M3 - Preprint

BT - Photon count estimation in single-molecule localization microscopy

ER -

Photon count estimation in single-molecule localization microscopy

Abstract

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