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PALM and STORM: Into large fields and high-throughput microscopy with sCMOS detectors

Almada, P; Culley, S; Henriques, R; (2015) PALM and STORM: Into large fields and high-throughput microscopy with sCMOS detectors. Methods , 88 pp. 109-121. 10.1016/j.ymeth.2015.06.004. Green open access

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Abstract

Single Molecule Localization Microscopy (SMLM) techniques such as Photo-Activation Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) enable fluorescence microscopy super-resolution: the overcoming of the resolution barrier imposed by the diffraction of light. These techniques are based on acquiring hundreds or thousands of images of single molecules, locating them and reconstructing a higher-resolution image from the high-precision localizations. These methods generally imply a considerable trade-off between imaging speed and resolution, limiting their applicability to high-throughput workflows. Recent advancements in scientific Complementary Metal-Oxide Semiconductor (sCMOS) camera sensors and localization algorithms reduce the temporal requirements for SMLM, pushing it toward high-throughput microscopy. Here we outline the decisions researchers face when considering how to adapt hardware on a new system for sCMOS sensors with high-throughput in mind.

Type: Article
Title: PALM and STORM: Into large fields and high-throughput microscopy with sCMOS detectors
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.ymeth.2015.06.004
Publisher version: http://dx.doi.org/10.1016/j.ymeth.2015.06.004
Language: English
Additional information: © 2015. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
Keywords: Hardware, Homogenization, Single-molecule localization, sCMOS
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL
URI: https://discovery.ucl.ac.uk/id/eprint/1469892
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