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25 February 2010 Super-resolved position and orientation estimation of fluorescent dipoles using 3-D steerable filters
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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.
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S. Geissbuehler, F. Aguet, I. Maerki, and T. Lasser "Super-resolved position and orientation estimation of fluorescent dipoles using 3-D steerable filters", Proc. SPIE 7571, Single Molecule Spectroscopy and Imaging III, 75710P (25 February 2010); doi: 10.1117/12.841783; https://doi.org/10.1117/12.841783
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