Designing the Focal Plane Spacing for Multifocal Plane Microscopy

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Designing the Focal Plane Spacing for Multifocal Plane Microscopy

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title Designing the Focal Plane Spacing for Multifocal Plane Microscopy
contributor.author Tahmasbi, Amir
contributor.author Ram, Sripad
contributor.author Chao, Jerry
contributor.author Abraham, Anish V.
contributor.author Tang, Felix W.
contributor.author Ward, E. S.
contributor.author Ober, Raimund J.
contributor.VIAF 279877162 (Ober, RJ)
description.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.
description.sponsorship US National Institutes of Health (R01 GM085575)
identifier.issn 1094-4087
identifier.uri http://hdl.handle.net/10735.1/4173
identifier.bibliographicCitation Tahmasbi, A., S. Ram, J. Chao, A. V. Abraham, et al. 2014. "Designing the focal plane spacing for multifocal plane microscopy." Optics Express 22(14): 16706-16721.
identifier.volume 22
identifier.issue 14
identifier.startpage 16706
subject Multifocal plane microscopy
subject Practical localization accuracy measure
subject Focal planes
date.issued 2014-06-30
publisher Optical Society of American (OSA)
relation.uri http://dx.doi.org/10.1364/OE.22.016706
rights ©2014 Optical Society of America
rights.holder Optical Society of America
language.iso en
source.journal Optics Express

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