Super-Resolved Position and Orientation Estimation of Fluorescent Dipoles Using 3-D Steerable Filters
Proceedings of SPIE, the International Society for Optical Engineering/Proceedings of SPIE(2010)
摘要
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.
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关键词
Super-resolution,dipole imaging,single molecule,orientation estimation,localization,steerable filters
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