Nguyen, D.T., Rahmani, A. orcid.org/0000-0002-2730-6099, Ponjavic, A. orcid.org/0000-0002-7561-1127 et al. (2 more authors) (2025) Active Remote Focus Stabilization in Oblique Plane Microscopy. Biomedical Optics Express, 16 (3). pp. 1216-1224. ISSN 2156-7085
Abstract
Light-sheet fluorescence microscopy (LSFM) has demonstrated great potential in the life sciences owing to its efficient volumetric imaging capabilities. For long-term imaging, the light-sheet typically needs to be stabilized to the detection focal plane for the best imaging results. Current light-sheet stabilization methods rely on fluorescence emission from the sample, which may interrupt scientific imaging and add to sample photobleaching. Here, we show that for oblique plane microscopes (OPM), a subset of LSFM where a single primary objective is used for illumination and detection, light-sheet stabilization can be achieved without expending sample fluorescence. Our method achieves ∼21 nm axial precision and maintains the light-sheet well within the depth of focus of the detection system for hour-long acquisition runs in a lab environment that would otherwise detune the system. We demonstrate subcellular imaging of the actin skeleton in melanoma cancer cells with a stabilized OPM.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2025 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Physics and Astronomy (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 11 Mar 2025 17:01 |
Last Modified: | 11 Mar 2025 17:01 |
Status: | Published |
Publisher: | Optica Publishing Group |
Identification Number: | 10.1364/boe.553545 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:224260 |