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

• DOI: 10.1117/1.nph.11.3.035006
• Publication Date: 2024-08-07T12:47:36Z
• Authors: Asaph Zylbertal; Isaac H. Bianco
• Keywords: Sample (material); Upgrade

SignificanceLight-sheet microscopy is a powerful imaging technique that achieves optical sectioning via selective illumination of individual sample planes. However, when the contains opaque or scattering tissues, incident light sheet may not be able to uniformly excite entire sample. For example, in context larval zebrafish whole-brain imaging, occlusion by eyes prevents access significant portion brain during common implementations using unidirectional illumination.AimWe introduce mirror-assisted light-sheet (mLSM), simple inexpensive method can implemented on existing digitally scanned setups adding single element—a mirrored micro-prism. The prism placed near generate second excitation path for accessing previously obstructed regions.ApproachScanning laser beam onto mirror generates second, reflected circumvents occlusion. Online tuning scanning and power waveforms enables uniform with dual illumination.ResultsmLSM produces cellular-resolution images regions inaccessible illumination. quality illuminated direct adjustable translating objective. does interfere visually guided behavior.ConclusionsmLSM presents an easy-to-implement, cost-effective way upgrade system obtain more data from biological