Super-resolved position and orientation estimation of fluorescent dipoles using 3-D steerable filters

S. Geissbuehler; F. Aguet; I. Maerki; T. Lasser

doi:10.1117/12.841783

25 February 2010 Super-resolved position and orientation estimation of fluorescent dipoles using 3-D steerable filters

S. Geissbuehler, F. Aguet, I. Maerki, T. Lasser

Proceedings Volume 7571, Single Molecule Spectroscopy and Imaging III; 75710P (2010) https://doi.org/10.1117/12.841783
Event: SPIE BiOS, 2010, San Francisco, California, United States

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.

Citation Download Citation

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); https://doi.org/10.1117/12.841783

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available