Dataset Information

Mirror-assisted light-sheet microscopy: a simple upgrade to enable bi-directional sample excitation.

ABSTRACT:

Significance

Light-sheet microscopy is a powerful imaging technique that achieves optical sectioning via selective illumination of individual sample planes. However, when the sample contains opaque or scattering tissues, the incident light sheet may not be able to uniformly excite the entire sample. For example, in the context of larval zebrafish whole-brain imaging, occlusion by the eyes prevents access to a significant portion of the brain during common implementations using unidirectional illumination.

Aim

We introduce mirror-assisted light-sheet microscopy (mLSM), a simple inexpensive method that can be implemented on existing digitally scanned light-sheet setups by adding a single optical element-a mirrored micro-prism. The prism is placed near the sample to generate a second excitation path for accessing previously obstructed regions.

Approach

Scanning the laser beam onto the mirror generates a second, reflected illumination path that circumvents the occlusion. Online tuning of the scanning and laser power waveforms enables near uniform sample excitation with dual illumination.

Results

mLSM produces cellular-resolution images of zebrafish brain regions inaccessible to unidirectional illumination. The imaging quality in regions illuminated by the direct or reflected sheet is adjustable by translating the excitation objective. The prism does not interfere with visually guided behavior.

Conclusions

mLSM presents an easy-to-implement, cost-effective way to upgrade an existing light-sheet system to obtain more imaging data from a biological sample.

SUBMITTER: Zylbertal A

PROVIDER: S-EPMC11304984 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature

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Publications

Mirror-assisted light-sheet microscopy: a simple upgrade to enable bi-directional sample excitation.

Zylbertal Asaph A Bianco Isaac H IH

Neurophotonics 20240701 3

<h4>Significance</h4>Light-sheet microscopy is a powerful imaging technique that achieves optical sectioning via selective illumination of individual sample planes. However, when the sample contains opaque or scattering tissues, the incident light sheet may not be able to uniformly excite the entire sample. For example, in the context of larval zebrafish whole-brain imaging, occlusion by the eyes prevents access to a significant portion of the brain during common implementations using unidirecti ...[more]

PMID: 39114857

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Dataset Information

Mirror-assisted light-sheet microscopy: a simple upgrade to enable bi-directional sample excitation.

Significance

Aim

Approach

Results

Conclusions

Publications

Mirror-assisted light-sheet microscopy: a simple upgrade to enable bi-directional sample excitation.

Similar Datasets

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