Super-Resolution Orientation Estimation and Localization of Fluorescent Dipoles Using 3-D Steerable Filters
F. Aguet, S. Geissbühler, I. Märki, T. Lasser, M. Unser
Optics Express, vol. 17, no. 8, pp. 6829–6848, April 9, 2009.
Fluorophores that are fixed during image acquisition produce a diffraction pattern that is characteristic of the orientation of the fluorophore's underlying dipole. Fluorescence localization microscopy techniques such as PALM and STORM achieve super-resolution by applying Gaussian-based fitting algorithms to in-focus images of individual fluorophores; when applied to fixed dipoles, this can lead to a bias in the range of 5-20 nm. We introduce a method for the joint estimation of position and orientation of dipoles, based on the representation of a physically realistic image formation model as a 3-D steerable filter. Our approach relies on a single, defocused acquisition. We establish theoretical, localization-based resolution limits on estimation accuracy using Cramér-Rao bounds, and experimentally show that estimation accuracies of at least 5 nm for position and of at least 2 degrees for orientation can be achieved. Patterns generated by applying the image formation model to estimated position/orientation pairs closely match experimental observations.
@ARTICLE(http://bigwww.epfl.ch/publications/aguet0901.html,
AUTHOR="Aguet, F. and Geissb{\"{u}}hler, S. and M{\"{a}}rki, I. and
Lasser, T. and Unser, M.",
TITLE="Super-Resolution Orientation Estimation and Localization of
Fluorescent Dipoles Using \mbox{3-D} Steerable Filters",
JOURNAL="Optics Express",
YEAR="2009",
volume="17",
number="8",
pages="6829--6848",
month="April 9,",
note="")