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31 July 2024 Posterior approach to correct for focal plane offsets in lattice light-sheet structured illumination microscopy
Yu Shi, Tim A. Daugird, Wesley R. Legant
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Journal of Biomedical Optics, Vol. 29, Issue 8, 086502 (July 2024). https://doi.org/10.1117/1.JBO.29.8.086502
Abstract

Significance

Lattice light-sheet structured illumination microscopy (latticeSIM) has proven highly effective in producing three-dimensional images with super resolution rapidly and with minimal photobleaching. However, due to the use of two separate objectives, sample-induced aberrations can result in an offset between the planes of excitation and detection, causing artifacts in the reconstructed images.

Aim

We introduce a posterior approach to detect and correct the axial offset between the excitation and detection focal planes in latticeSIM and provide a method to minimize artifacts in the reconstructed images.

Approach

We utilized the residual phase information within the overlap regions of the laterally shifted structured illumination microscopy information components in frequency space to retrieve the axial offset between the excitation and the detection focal planes in latticeSIM.

Results

We validated our technique through simulations and experiments, encompassing a range of samples from fluorescent beads to subcellular structures of adherent cells. We also show that using transfer functions with the same axial offset as the one present during data acquisition results in reconstructed images with minimal artifacts and salvages otherwise unusable data.

Conclusion

We envision that our method will be a valuable addition to restore image quality in latticeSIM datasets even for those acquired under non-ideal experimental conditions.
CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
Citation Download Citation
Yu Shi, Tim A. Daugird, and Wesley R. Legant "Posterior approach to correct for focal plane offsets in lattice light-sheet structured illumination microscopy," Journal of Biomedical Optics 29(8), 086502 (31 July 2024). https://doi.org/10.1117/1.JBO.29.8.086502
Received: 18 April 2024; Accepted: 24 June 2024; Published: 31 July 2024
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KEYWORDS
Image restoration
Optical transfer functions
Light sources and illumination
Point spread functions
Hydrogels
Biological samples
Object detection
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