Computational localization microscopy with extended axial range

Zhou, Y. , Zammit, P., Carles Santacana, G. and Harvey, A. R. (2018) Computational localization microscopy with extended axial range. Optics Express, 26(2), pp. 7563-7577. (doi: 10.1364/OE.26.007563) (PMID:29609310)

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Abstract

A new single-aperture 3D particle-localization and tracking technique is presented that demonstrates an increase in depth range by more than an order of magnitude without compromising optical resolution and throughput. We exploit the extended depth range and depth-dependent translation of an Airy-beam PSF for 3D localization over an extended volume in a single snapshot. The technique is applicable to all bright-field and fluorescence modalities for particle localization and tracking, ranging from super-resolution microscopy through to the tracking of fluorescent beads and endogenous particles within cells. We demonstrate and validate its application to real-time 3D velocity imaging of fluid flow in capillaries using fluorescent tracer beads. An axial localization precision of 50 nm was obtained over a depth range of 120μm using a 0.4NA, 20× microscope objective. We believe this to be the highest ratio of axial range-to-precision reported to date.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Carles Santacana, Dr Guillem and Zhou, Mr Yongzhuang and Zammit, Mr Paul and Harvey, Professor Andy
Authors: Zhou, Y., Zammit, P., Carles Santacana, G., and Harvey, A. R.
College/School:College of Science and Engineering > School of Physics and Astronomy
Journal Name:Optics Express
Publisher:Optical Society of America
ISSN:1094-4087
ISSN (Online):1094-4087
Published Online:15 March 2018
Copyright Holders:Copyright © 2018 The Optical Society of America
First Published:First published in Optics Express 26(6):7563-7577
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
608951Engineering and Physical Sciences Doctoral Training Grant 2012-16Mary Beth KneafseyEngineering and Physical Sciences Research Council (EPSRC)EP/K503058/1VPO VICE PRINCIPAL RESEARCH & ENTERPRISE
729121Seeing within extended volumes and 3D structures: 3D imaging and applicationsGuillem Carles SantacanaLeverhulme Trust (LEVERHUL)ECF-2016-757S&E P&A - PHYSICS & ASTRONOMY