An information-theoretic approach to designing the plane spacing for multifocal plane microscopy
An information-theoretic approach to designing the plane spacing for multifocal plane microscopy
Multifocal plane microscopy (MUM) is a 3D imaging modality which enables the localization and tracking of single molecules at high spatial and temporal resolution by simultaneously imaging distinct focal planes within the sample. MUM overcomes the depth discrimination problem of conventional microscopy and allows high accuracy localization of a single molecule in 3D along the z-axis. An important question in the design of MUM experiments concerns the appropriate number of focal planes and their spacings to achieve the best possible 3D localization accuracy along the z-axis. Ideally, it is desired to obtain a 3D localization accuracy that is uniform over a large depth and has small numerical values, which guarantee that the single molecule is continuously detectable. Here, we address this concern by developing a plane spacing design strategy based on the Fisher information. In particular, we analyze the Fisher information matrix for the 3D localization problem along the z-axis and propose spacing scenarios termed the strong coupling and the weak coupling spacings, which provide appropriate 3D localization accuracies. Using these spacing scenarios, we investigate the detectability of the single molecule along the z-axis and study the effect of changing the number of focal planes on the 3D localization accuracy. We further review a software module we recently introduced, the MUMDesignTool, that helps to design the plane spacings for a MUM setup.
Cramér-Rao lower bound, Fisher Information Matrix, Single molecule microscopy
Tahmasbi, Amir
5a377bd3-4578-434f-919a-87804245791e
Ram, Sripad
559bd560-3817-4e53-8c7a-2f08e4518412
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Abraham, Anish V.
4f71ee5b-b0b1-4e0e-8f64-5cea9ae8f0e0
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
2015
Tahmasbi, Amir
5a377bd3-4578-434f-919a-87804245791e
Ram, Sripad
559bd560-3817-4e53-8c7a-2f08e4518412
Chao, Jerry
550e20b0-8365-42e3-a6fc-1048eb8c2e47
Abraham, Anish V.
4f71ee5b-b0b1-4e0e-8f64-5cea9ae8f0e0
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Tahmasbi, Amir, Ram, Sripad, Chao, Jerry, Abraham, Anish V., Ward, E. Sally and Ober, Raimund J.
(2015)
An information-theoretic approach to designing the plane spacing for multifocal plane microscopy.
In Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII.
vol. 9330,
SPIE..
(doi:10.1117/12.2076769).
Record type:
Conference or Workshop Item
(Paper)
Abstract
Multifocal plane microscopy (MUM) is a 3D imaging modality which enables the localization and tracking of single molecules at high spatial and temporal resolution by simultaneously imaging distinct focal planes within the sample. MUM overcomes the depth discrimination problem of conventional microscopy and allows high accuracy localization of a single molecule in 3D along the z-axis. An important question in the design of MUM experiments concerns the appropriate number of focal planes and their spacings to achieve the best possible 3D localization accuracy along the z-axis. Ideally, it is desired to obtain a 3D localization accuracy that is uniform over a large depth and has small numerical values, which guarantee that the single molecule is continuously detectable. Here, we address this concern by developing a plane spacing design strategy based on the Fisher information. In particular, we analyze the Fisher information matrix for the 3D localization problem along the z-axis and propose spacing scenarios termed the strong coupling and the weak coupling spacings, which provide appropriate 3D localization accuracies. Using these spacing scenarios, we investigate the detectability of the single molecule along the z-axis and study the effect of changing the number of focal planes on the 3D localization accuracy. We further review a software module we recently introduced, the MUMDesignTool, that helps to design the plane spacings for a MUM setup.
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e-pub ahead of print date: 9 March 2015
Published date: 2015
Venue - Dates:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII, , San Francisco, United States, 2015-02-09 - 2015-02-12
Keywords:
Cramér-Rao lower bound, Fisher Information Matrix, Single molecule microscopy
Identifiers
Local EPrints ID: 423667
URI: http://eprints.soton.ac.uk/id/eprint/423667
PURE UUID: d45f3ea4-8b0b-48a3-a8db-1f89755739ea
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Date deposited: 27 Sep 2018 16:30
Last modified: 16 Mar 2024 04:37
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Contributors
Author:
Amir Tahmasbi
Author:
Sripad Ram
Author:
Jerry Chao
Author:
Anish V. Abraham
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