Cleared tissue dual-view oblique plane microscopy
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Published version
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Author(s)
Type
Journal Article
Abstract
We present a dual-view oblique plane microscope (dOPM) for imaging thick optically cleared tissue samples using a silicone immersion primary objective. The custom-designed remote refocusing relay utilizes stock optics to achieve remote refocusing in refractive index-matched samples. The spatial resolution of the system was characterized using a series of fluorescent bead phantoms with refractive indices ranging from 1.4-1.5, with the point spread function full width at half maximum measuring ∼0.5 µm laterally and ∼1 µm axially for a refractive index-matched bead sample, with minimal degradation over a > 250 µm axial range. We characterize how the remote refocusing performance at sample refractive indices up to n = 1.5 can be partially compensated for using adjustment of the correction collar on the primary objective. We apply the system to imaging a range of biological samples with varied refractive indices. Combined with tiled acquisition, image stitching, and multi-view image fusion, the microscope enables multicolour imaging of mm-wide and ∼250 µm thick optically cleared mouse ovarian cancer, bone marrow, and colon tissue samples, and a whole Drosophila melanogaster fruit fly brain. The system offers a platform for fast and high-resolution, multicolour volumetric imaging across spatial scales, integrated on a commercially available microscope frame.
Date Issued
2025-09-01
Date Acceptance
2025-08-12
Citation
Biomedical Optics Express, 2025, 16 (9), pp.3725-3739
ISSN
2156-7085
Publisher
Optical Society of America (OSA)
Start Page
3725
End Page
3739
Journal / Book Title
Biomedical Optics Express
Volume
16
Issue
9
Copyright Statement
© 2025 The Author(s). Published by Optica Publishing Group under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.
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Publication Status
Published