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Accuracy of the gaussian point spread function model in 2D localization microscopy.

Sjoerd Stallinga1, Bernd Rieger

  • 1Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands. s.stallinga@tudelft.nl

Optics Express
|December 18, 2010
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Summary
This summary is machine-generated.

The gaussian Point Spread Function (PSF) model is inaccurate for fixed dipole emitters in super-resolution microscopy. Optical aberrations and dipole orientation cause significant localization errors, necessitating advanced PSF models.

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Area of Science:

  • Optical microscopy
  • Super-resolution imaging
  • Biophysics

Background:

  • The Gaussian function is a common Point Spread Function (PSF) model in localization microscopy due to its simplicity.
  • However, its optical basis is questionable, potentially impacting localization accuracy.

Purpose of the Study:

  • To investigate the impact of emission dipole orientation and optical aberrations on localization accuracy using a Gaussian PSF model.
  • To determine the necessity of advanced PSF models for precise emitter localization.

Main Methods:

  • Simulated image spots incorporating high numerical aperture, media interfaces, polarization, dipole orientation, and aberrations.
  • Fitting simulated spots using a Maximum Likelihood Estimator with a Gaussian PSF model.

Main Results:

  • For freely rotating dipoles, the Gaussian PSF model yields optimal localization accuracy, even with aberrations within the Marechal diffraction limit.
  • For fixed dipole orientations, localization errors up to 40 nm occur, independent of photon count, indicating systematic bias.

Conclusions:

  • The Gaussian PSF model is inappropriate for emitters with fixed dipole orientations.
  • Advanced, non-Gaussian PSF models are essential for accurate localization in super-resolution microscopy under realistic optical conditions.