Three-dimensional single molecule localization microscopy relies on the fitting of the individual molecules with a point spread function (PSF) model. The reconstructed images often show local squeezing or expansion in z. A common cause is depth-induced aberrations in conjunction with an imperfect PSF model calibrated from beads on a coverslip, resulting in a mismatch between measured PSF and real PSF. Here, we developed a strategy for accurate z-localization in which we use the imperfect PSF model for fitting, determine the fitting errors and correct for them in a post-processing step. We present an open-source software tool and a simple experimental calibration procedure that allow retrieving accurate z-positions in any PSF engineering approach or fitting modality, even at large imaging depths.
:C$\backslash$:/Users/Karoline/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/Li et al. - 2019 - Depth-dependent PSF calibration and aberration correction for 3D single-molecule localization.pdf:pdf
%0 Journal Article
%1 Li2019
%A Li, Yiming
%A Wu, Yu-Le
%A Hoess, Philipp
%A Mund, Markus
%A Ries, Jonas
%D 2019
%I The Optical Society
%J Biomedical Optics Express
%K 3D Aberration aperture correction internal microscopy numerical psf reflection software
%N 6
%P 2708
%R 10.1364/boe.10.002708
%T Depth-dependent PSF calibration and aberration correction for 3D single-molecule localization
%V 10
%X Three-dimensional single molecule localization microscopy relies on the fitting of the individual molecules with a point spread function (PSF) model. The reconstructed images often show local squeezing or expansion in z. A common cause is depth-induced aberrations in conjunction with an imperfect PSF model calibrated from beads on a coverslip, resulting in a mismatch between measured PSF and real PSF. Here, we developed a strategy for accurate z-localization in which we use the imperfect PSF model for fitting, determine the fitting errors and correct for them in a post-processing step. We present an open-source software tool and a simple experimental calibration procedure that allow retrieving accurate z-positions in any PSF engineering approach or fitting modality, even at large imaging depths.
@article{Li2019,
abstract = {Three-dimensional single molecule localization microscopy relies on the fitting of the individual molecules with a point spread function (PSF) model. The reconstructed images often show local squeezing or expansion in z. A common cause is depth-induced aberrations in conjunction with an imperfect PSF model calibrated from beads on a coverslip, resulting in a mismatch between measured PSF and real PSF. Here, we developed a strategy for accurate z-localization in which we use the imperfect PSF model for fitting, determine the fitting errors and correct for them in a post-processing step. We present an open-source software tool and a simple experimental calibration procedure that allow retrieving accurate z-positions in any PSF engineering approach or fitting modality, even at large imaging depths.},
added-at = {2020-03-23T21:12:34.000+0100},
author = {Li, Yiming and Wu, Yu-Le and Hoess, Philipp and Mund, Markus and Ries, Jonas},
biburl = {https://www.bibsonomy.org/bibtex/2eb6d8a0dbd3e846305e031ff033e895b/kfriedl},
doi = {10.1364/boe.10.002708},
file = {:C$\backslash$:/Users/Karoline/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/Li et al. - 2019 - Depth-dependent PSF calibration and aberration correction for 3D single-molecule localization.pdf:pdf},
interhash = {46dfb22c0225a8ab61dbe9ad42e6bfd2},
intrahash = {eb6d8a0dbd3e846305e031ff033e895b},
issn = {2156-7085},
journal = {Biomedical Optics Express},
keywords = {3D Aberration aperture correction internal microscopy numerical psf reflection software},
month = jun,
number = 6,
pages = 2708,
publisher = {The Optical Society},
timestamp = {2020-03-23T22:27:15.000+0100},
title = {{Depth-dependent PSF calibration and aberration correction for 3D single-molecule localization}},
volume = 10,
year = 2019
}