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A comparative study of high resolution microscopy imaging modalities using a three-dimensional resolution measure

A comparative study of high resolution microscopy imaging modalities using a three-dimensional resolution measure
A comparative study of high resolution microscopy imaging modalities using a three-dimensional resolution measure

From an acquired image, single molecule microscopy makes possible the determination of the distance separating two closely spaced biomolecules in three-dimensional (3D) space. Such distance information can be an important indicator of the nature of the biomolecular interaction. Distance determination, however, is especially difficult when, for example, the imaged point sources are very close to each other or are located near the focal plane of the imaging setup. In the context of such challenges, we compare the limits of the distance estimation accuracy for several high resolution 3D imaging modalities. The comparisons are made using a Cramer-Rao lower bound-based 3D resolution measure which predicts the best possible accuracy with which a given distance can be estimated. Modalities which separate the detection of individual point sources (e.g., using photoactivatable fluorophores) are shown to provide the best accuracy limits when the two point sources are very close to each other and/or are oriented near parallel to the optical axis. Meanwhile, modalities which implement the simultaneous imaging of the point sources from multiple focal planes perform best when given a near-focus point source pair. We also demonstrate that the maximum likelihood estimator is capable of attaining the limit of the accuracy predicted for each modality.

1094-4087
24377-24402
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Ram, Sripad
559bd560-3817-4e53-8c7a-2f08e4518412
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
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36

Chao, Jerry, Ram, Sripad, Ward, E. Sally and Ober, Raimund J. (2009) A comparative study of high resolution microscopy imaging modalities using a three-dimensional resolution measure. Optics Express, 17 (26), 24377-24402. (doi:10.1364/OE.17.024377).

Record type: Article

Abstract

From an acquired image, single molecule microscopy makes possible the determination of the distance separating two closely spaced biomolecules in three-dimensional (3D) space. Such distance information can be an important indicator of the nature of the biomolecular interaction. Distance determination, however, is especially difficult when, for example, the imaged point sources are very close to each other or are located near the focal plane of the imaging setup. In the context of such challenges, we compare the limits of the distance estimation accuracy for several high resolution 3D imaging modalities. The comparisons are made using a Cramer-Rao lower bound-based 3D resolution measure which predicts the best possible accuracy with which a given distance can be estimated. Modalities which separate the detection of individual point sources (e.g., using photoactivatable fluorophores) are shown to provide the best accuracy limits when the two point sources are very close to each other and/or are oriented near parallel to the optical axis. Meanwhile, modalities which implement the simultaneous imaging of the point sources from multiple focal planes perform best when given a near-focus point source pair. We also demonstrate that the maximum likelihood estimator is capable of attaining the limit of the accuracy predicted for each modality.

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More information

Accepted/In Press date: 17 December 2009
e-pub ahead of print date: 18 December 2009
Published date: 21 December 2009

Identifiers

Local EPrints ID: 423605
URI: https://eprints.soton.ac.uk/id/eprint/423605
ISSN: 1094-4087
PURE UUID: 600bc48a-2de5-4203-9d08-c8e01e32155a
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

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Contributors

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

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