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Digital holography-based 3D particle localization for single-molecule tweezer techniques

Flewellen, James L; Minoughan, Sophie; Garcia, Isabel Llorente; Tolar, Pavel; (2022) Digital holography-based 3D particle localization for single-molecule tweezer techniques. Biophysical Journal 10.1016/j.bpj.2022.06.001. (In press). Green open access

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Abstract

We present a three-dimensional (3D) imaging technique for the fast tracking of microscopic objects in a fluid environment. Our technique couples digital holographic microscopy with three-dimensional localization via parabolic masking. Compared with existing approaches, our method reconstructs 3D volumes from single-plane images, which greatly simplifies image acquisition, reduces the demand on microscope hardware, and facilitates tracking higher densities of microscopic particles while maintaining similar levels of precision. We demonstrate utility of this method in magnetic tweezer experiments, opening their use to multiplexed single-molecule force spectroscopy assays, which were previously limited by particle crowding and fast dissociation times. We propose that our technique will also be useful in other applications that involve the tracking of microscopic objects in three dimensions, such as studies of microorganism motility and 3D flow characterization of microfluidic devices.

Type: Article
Title: Digital holography-based 3D particle localization for single-molecule tweezer techniques
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.bpj.2022.06.001
Publisher version: https://doi.org/10.1016/j.bpj.2022.06.001
Language: English
Additional information: © 2022 Published by Biophysical Society under a Creative Commons license (https://creativecommons.org/licenses/by/4.0/).
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL
URI: https://discovery.ucl.ac.uk/id/eprint/10150599
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