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Proceedings Paper

Super-resolved position and orientation estimation of fluorescent dipoles using 3-D steerable filters
Author(s): S. Geissbuehler; F. Aguet; I. Maerki; T. Lasser
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Paper Abstract

The diffraction patterns of fixed fluorophores are characteristic of the orientation of the molecules' underlying dipole. Fluorescence localization microscopy techniques such as PALM and STORM achieve super-resolution by sequentially imaging sparse subsets of fluorophores, which are localized by means of Gaussian-based localization. This approach is based on the assumption of isotropic emitters, where the diffraction pattern corresponds to a section of the point spread function. Applied to fixed fluorophores, it can lead to an estimation bias in the range of 5-20nm. We introduce a method for the joint estimation of position and orientation of single fluorophores, based on an accurate image formation model expressed as a 3-D steerable filter. We demonstrate experimental estimation accuracies of 5 nm for position and 2 degrees for orientation.

Paper Details

Date Published: 25 February 2010
PDF: 7 pages
Proc. SPIE 7571, Single Molecule Spectroscopy and Imaging III, 75710P (25 February 2010); doi: 10.1117/12.841783
Show Author Affiliations
S. Geissbuehler, École Polytechnique Fédérale de Lausanne (Switzerland)
F. Aguet, École Polytechnique Fédérale de Lausanne (Switzerland)
I. Maerki, École Polytechnique Fédérale de Lausanne (Switzerland)
T. Lasser, École Polytechnique Fédérale de Lausanne (Switzerland)

Published in SPIE Proceedings Vol. 7571:
Single Molecule Spectroscopy and Imaging III
Jörg Enderlein; Zygmunt Karol Gryczynski; Rainer Erdmann, Editor(s)

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