The University of Southampton
University of Southampton Institutional Repository

Investigation of the numerics of point spread function integration in single molecule localization

Investigation of the numerics of point spread function integration in single molecule localization
Investigation of the numerics of point spread function integration in single molecule localization

The computation of point spread functions, which are typically used to model the image profile of a single molecule, represents a central task in the analysis of single molecule microscopy data. To determine how the accuracy of the computation affects how well a single molecule can be localized, we investigate how the fineness with which the point spread function is integrated over an image pixel impacts the performance of the maximum likelihood location estimator. We consider both the Airy and the two-dimensional Gaussian point spread functions. Our results show that the point spread function needs to be adequately integrated over a pixel to ensure that the estimator closely recovers the true location of the single molecule with an accuracy that is comparable to the best possible accuracy as determined using the Fisher information formalism. Importantly, if integration with an insufficiently fine step size is carried out, the resulting estimates can be significantly different from the true location, particularly when the image data is acquired at relatively low magnifications. We also present a methodology for determining an adequate step size for integrating the point spread function.

1094-4087
16866-16883
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Ram, Sripad
559bd560-3817-4e53-8c7a-2f08e4518412
Lee, Taiyoon
763ca47b-a0fd-43e9-9604-ea148726fa6e
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Ram, Sripad
559bd560-3817-4e53-8c7a-2f08e4518412
Lee, Taiyoon
763ca47b-a0fd-43e9-9604-ea148726fa6e
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36

Chao, Jerry, Ram, Sripad, Lee, Taiyoon, Ward, E. Sally and Ober, Raimund J. (2015) Investigation of the numerics of point spread function integration in single molecule localization. Optics Express, 23 (13), 16866-16883. (doi:10.1364/OE.23.016866).

Record type: Article

Abstract

The computation of point spread functions, which are typically used to model the image profile of a single molecule, represents a central task in the analysis of single molecule microscopy data. To determine how the accuracy of the computation affects how well a single molecule can be localized, we investigate how the fineness with which the point spread function is integrated over an image pixel impacts the performance of the maximum likelihood location estimator. We consider both the Airy and the two-dimensional Gaussian point spread functions. Our results show that the point spread function needs to be adequately integrated over a pixel to ensure that the estimator closely recovers the true location of the single molecule with an accuracy that is comparable to the best possible accuracy as determined using the Fisher information formalism. Importantly, if integration with an insufficiently fine step size is carried out, the resulting estimates can be significantly different from the true location, particularly when the image data is acquired at relatively low magnifications. We also present a methodology for determining an adequate step size for integrating the point spread function.

Full text not available from this repository.

More information

Accepted/In Press date: 28 May 2015
e-pub ahead of print date: 18 June 2015
Published date: 29 June 2015

Identifiers

Local EPrints ID: 423660
URI: https://eprints.soton.ac.uk/id/eprint/423660
ISSN: 1094-4087
PURE UUID: 1277d268-09d2-4fc9-b52c-7a45782f54c9
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

Catalogue record

Date deposited: 27 Sep 2018 16:30
Last modified: 20 Jul 2019 00:22

Export record

Altmetrics

Contributors

Author: Jerry Chao
Author: Sripad Ram
Author: Taiyoon Lee
Author: E. Sally Ward ORCID iD
Author: Raimund J. Ober ORCID iD

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×