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

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.

Published in:
Proceedings of SPIE, 7571, 75710P
Presented at:
BiOS 2010 - Single Molecule Spectroscopy and Imaging III, San Francisco, January 23-28, 2010
Spie-Int Soc Optical Engineering, Po Box 10, Bellingham, Wa 98227-0010 Usa
Invited Paper

 Record created 2010-03-12, last modified 2018-09-13

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