Mesoscale standing wave imaging

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Foylan, Shannan and Schniete, Jana Katharina and Kölln, Lisa Sophie and Dempster, John and Hansen, Carsten Gram and Shaw, Michael and Bushell, Trevor John and McConnell, Gail (2023) Mesoscale standing wave imaging. Journal of Microscopy, 295 (1). pp. 33-41. ISSN 0022-2720 (https://doi.org/10.1111/jmi.13189)

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Abstract

Standing wave (SW) microscopy is a method that uses an interference pattern to excite fluorescence from labelled cellular structures and produces high-resolution images of three-dimensional objects in a two-dimensional dataset. SW microscopy is performed with high-magnification, high-numerical aperture objective lenses, and while this results in high-resolution images, the field of view is very small. Here we report upscaling of this interference imaging method from the microscale to the mesoscale using the Mesolens, which has the unusual combination of a low-magnification and high-numerical aperture. With this method, we produce SW images within a field of view of 4.4 mm × 3.0 mm that can readily accommodate over 16,000 cells in a single dataset. We demonstrate the method using both single-wavelength excitation and the multi-wavelength SW method TartanSW. We show application of the method for imaging of fixed and living cells specimens, with the first application of SW imaging to study cells under flow conditions.

ORCID iDs

Foylan, Shannan, Schniete, Jana Katharina ORCID logoORCID: https://orcid.org/0000-0003-4274-5839, Kölln, Lisa Sophie, Dempster, John ORCID logoORCID: https://orcid.org/0000-0003-2199-2945, Hansen, Carsten Gram, Shaw, Michael, Bushell, Trevor John ORCID logoORCID: https://orcid.org/0000-0003-4145-9670 and McConnell, Gail ORCID logoORCID: https://orcid.org/0000-0002-7213-0686;
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