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research article

Super-resolution orientation estimation and localization of fluorescent dipoles using 3-D steerable filters

Aguet, François  
•
Geissbühler, Stefan  
•
Märki, Iwan  
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2009
Optics Express

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.

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Type
research article
DOI
10.1364/OE.17.006829
Web of Science ID

WOS:000265108900102

Author(s)
Aguet, François  
Geissbühler, Stefan  
Märki, Iwan  
Lasser, Theo  
Unser, Michael  
Date Issued

2009

Publisher

OSA

Published in
Optics Express
Volume

17

Issue

8

Start page

6829

End page

6848

Subjects

Fluorescence microscopy

•

Superresolution

•

Pattern recognition

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

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Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/40122
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