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Dual-view oblique plane microscopy (dOPM)

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journal contribution
posted on 16.12.2020, 09:20 by H Sparks, L Dent, C Bakal, A Behrens, G Salbreux, C Dunsby
© 2020 OSA - The Optical Society. All rights reserved. We present a new folded dual-view oblique plane microscopy (OPM) technique termed dOPM that enables two orthogonal views of the sample to be obtained by translating a pair of tilted mirrors in refocussing space. Using a water immersion 40× 1.15 NA primary objective, deconvolved image volumes of 200 nm beads were measured to have full width at half maxima (FWHM) of 0.35 ± 0.04 µm and 0.39 ± 0.02 µm laterally and 0.81 ± 0.07 µm axially. The measured z-sectioning value was 1.33 ± 0.45 µm using light-sheet FWHM in the frames of the two views of 4.99 ± 0.58 µm and 4.89 ± 0.63 µm. To qualitatively demonstrate that the system can reduce shadow artefacts while providing a more isotropic resolution, a multi-cellular spheroid approximately 100 µm in diameter was imaged.

Funding

Crick (Grant ID: 10039, Grant title: Behrens FC001039) Crick (Grant ID: 10317, Grant title: Salbreux FC001317) Engineering and Physical Sciences Research Council (Grant ID: EP/T003103/1, Grant title: EPSRC EP/T003103/1)

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