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. 2024 Jul;295(1):33-41.
doi: 10.1111/jmi.13189. Epub 2023 May 17.

Mesoscale standing wave imaging

Shannan Foylan  1 , Jana Katharina Schniete  1 , Lisa Sophie Kölln  1 , John Dempster  1 , Carsten Gram Hansen  2 , Michael Shaw  3   4 , Trevor John Bushell  1 , Gail McConnell  1
Affiliations
Free article

Mesoscale standing wave imaging

Shannan Foylan et al. J Microsc. 2024 Jul.
Free article

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.

Keywords: fluorescence; interference; mesoscopy; microscopy; standing wave; topography.

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