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  4. Super-resolved position and orientation from defocused images of fluorescent dipoles
 
conference presentation

Super-resolved position and orientation from defocused images of fluorescent dipoles

Geissbühler, Stefan  
•
Aguet, François
•
Märki, Iwan  
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2009
European Conference on Biomedical Optics

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 Cram´er-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
conference presentation
Author(s)
Geissbühler, Stefan  
Aguet, François
Märki, Iwan  
Lasser, Theo  
Unser, Michael
Date Issued

2009

Subjects

Fluorescence microscopy

•

Superresolution

•

Pattern recognition

URL

URL

http://www.osa.org/meetings/topicalmeetings/ecbo/
Written at

EPFL

EPFL units
LOB  
Event nameEvent placeEvent date
European Conference on Biomedical Optics

Munich

June 14–18 2009

Available on Infoscience
December 3, 2009
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/44588
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