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A state space based approach to localizing single molecules from multi-emitter images

A state space based approach to localizing single molecules from multi-emitter images
A state space based approach to localizing single molecules from multi-emitter images

Single molecule super-resolution microscopy is a powerful tool that enables imaging at sub-diffraction-limit resolution. In this technique, subsets of stochastically photoactivated fluorophores are imaged over a sequence of frames and accurately localized, and the estimated locations are used to construct a high-resolution image of the cellular structures labeled by the fluorophores. Available localization methods typically first determine the regions of the image that contain emitting fluorophores through a process referred to as detection. Then, the locations of the fluorophores are estimated accurately in an estimation step. We propose a novel localization method which combines the detection and estimation steps. The method models the given image as the frequency response of a multi-order system obtained with a balanced state space realization algorithm based on the singular value decomposition of a Hankel matrix, and determines the locations of intensity peaks in the image as the pole locations of the resulting system. The locations of the most significant peaks correspond to the locations of single molecules in the original image. Although the accuracy of the location estimates is reasonably good, we demonstrate that, by using the estimates as the initial conditions for a maximum likelihood estimator, refined estimates can be obtained that have a standard deviation close to the Cramér-Rao lower bound-based limit of accuracy. We validate our method using both simulated and experimental multi-emitter images.

Fluorescence microscopy, frequency response, multi-emitter localization, single molecule localization, single molecule microscopy, singular value decomposition, state space realization, super-resolution microscopy
SPIE
Vahid, Milad R.
6a1a88a4-9fcc-4ac5-84a4-0d4ee1088cc6
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Vahid, Milad R.
6a1a88a4-9fcc-4ac5-84a4-0d4ee1088cc6
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36

Vahid, Milad R., Chao, Jerry, Ward, E. Sally and Ober, Raimund J. (2017) A state space based approach to localizing single molecules from multi-emitter images. In Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV. vol. 10070, SPIE.. (doi:10.1117/12.2253175).

Record type: Conference or Workshop Item (Paper)

Abstract

Single molecule super-resolution microscopy is a powerful tool that enables imaging at sub-diffraction-limit resolution. In this technique, subsets of stochastically photoactivated fluorophores are imaged over a sequence of frames and accurately localized, and the estimated locations are used to construct a high-resolution image of the cellular structures labeled by the fluorophores. Available localization methods typically first determine the regions of the image that contain emitting fluorophores through a process referred to as detection. Then, the locations of the fluorophores are estimated accurately in an estimation step. We propose a novel localization method which combines the detection and estimation steps. The method models the given image as the frequency response of a multi-order system obtained with a balanced state space realization algorithm based on the singular value decomposition of a Hankel matrix, and determines the locations of intensity peaks in the image as the pole locations of the resulting system. The locations of the most significant peaks correspond to the locations of single molecules in the original image. Although the accuracy of the location estimates is reasonably good, we demonstrate that, by using the estimates as the initial conditions for a maximum likelihood estimator, refined estimates can be obtained that have a standard deviation close to the Cramér-Rao lower bound-based limit of accuracy. We validate our method using both simulated and experimental multi-emitter images.

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More information

Published date: 17 February 2017
Venue - Dates: Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV 2017, , San Francisco, United States, 2017-01-30 - 2017-02-01
Keywords: Fluorescence microscopy, frequency response, multi-emitter localization, single molecule localization, single molecule microscopy, singular value decomposition, state space realization, super-resolution microscopy

Identifiers

Local EPrints ID: 423678
URI: http://eprints.soton.ac.uk/id/eprint/423678
PURE UUID: f0ce6321-74b4-403b-92d4-3861214c608a
ORCID for E. Sally Ward: ORCID iD orcid.org/0000-0003-3232-7238
ORCID for Raimund J. Ober: ORCID iD orcid.org/0000-0002-1290-7430

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Date deposited: 27 Sep 2018 16:30
Last modified: 18 Mar 2024 03:48

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Contributors

Author: Milad R. Vahid
Author: Jerry Chao
Author: E. Sally Ward ORCID iD
Author: Raimund J. Ober ORCID iD

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