Designing the focal plane spacing for multifocal plane microscopy
Designing the focal plane spacing for multifocal plane microscopy
Multifocal plane microscopy (MUM) has made it possible to study subcellular dynamics in 3D at high temporal and spatial resolution by simultaneously imaging distinct planes within the specimen. MUM allows high accuracy localization of a point source along the z-axis since it overcomes the depth discrimination problem of conventional single plane microscopy. An important question in MUM experiments is how the number of focal planes and their spacings should be chosen to achieve the best possible localization accuracy along the z-axis. Here, we propose approaches based on the Fisher information matrix and report spacing scenarios called strong coupling and weak coupling which yield an appropriate 3D localization accuracy. We examine the effect of numerical aperture, magnification, photon count, emission wavelength and extraneous noise on the spacing scenarios. In addition, we investigate the effect of changing the number of focal planes on the 3D localization accuracy. We also introduce a new software package that provides a user-friendly framework to find appropriate plane spacings for a MUM setup. These developments should assist in optimizing MUM experiments.
16706-16721
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
Tang, Felix W.
fefc2616-fcda-4165-ac93-7af9007c666d
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
2014
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
Tang, Felix W.
fefc2616-fcda-4165-ac93-7af9007c666d
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., Tang, Felix W., Ward, E. Sally and Ober, Raimund J.
(2014)
Designing the focal plane spacing for multifocal plane microscopy.
Optics Express, 22 (14), .
(doi:10.1364/OE.22.016706).
Abstract
Multifocal plane microscopy (MUM) has made it possible to study subcellular dynamics in 3D at high temporal and spatial resolution by simultaneously imaging distinct planes within the specimen. MUM allows high accuracy localization of a point source along the z-axis since it overcomes the depth discrimination problem of conventional single plane microscopy. An important question in MUM experiments is how the number of focal planes and their spacings should be chosen to achieve the best possible localization accuracy along the z-axis. Here, we propose approaches based on the Fisher information matrix and report spacing scenarios called strong coupling and weak coupling which yield an appropriate 3D localization accuracy. We examine the effect of numerical aperture, magnification, photon count, emission wavelength and extraneous noise on the spacing scenarios. In addition, we investigate the effect of changing the number of focal planes on the 3D localization accuracy. We also introduce a new software package that provides a user-friendly framework to find appropriate plane spacings for a MUM setup. These developments should assist in optimizing MUM experiments.
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Accepted/In Press date: 20 June 2014
e-pub ahead of print date: 30 June 2014
Published date: 2014
Identifiers
Local EPrints ID: 423651
URI: http://eprints.soton.ac.uk/id/eprint/423651
ISSN: 1094-4087
PURE UUID: 77b23d17-8579-45dc-8aaf-371619e178ca
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Date deposited: 27 Sep 2018 16:30
Last modified: 16 Mar 2024 04:37
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Author:
Amir Tahmasbi
Author:
Sripad Ram
Author:
Jerry Chao
Author:
Anish V. Abraham
Author:
Felix W. Tang
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