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Updated: Jun 23, 2026

Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples
Published on: March 26, 2021
Scanning Bessel beam microscopy with a protected and corrective objective for solvent-cleared large samples
Chia-Ming Lee1, Po-Yen Lin2, Yu-Ting Tseng1
1Research Center for Applied Sciences, Academia Sinica, Taipei 11529, Taiwan.
None:
Emerging imaging strategies, such as tissue clearing and advanced light-sheet modalities, are transforming the visualization of cell differentiation, tissue reorganization, and morphogenesis in both model and non-model organisms, thereby driving future discoveries of conserved developmental programs and species-specific innovations. In this study, we present a versatile Bessel light sheet microscopy (BLX) system compatible with both immersion and capped air objectives, optimized for whole-mount three-dimensional imaging of cleared tissues. We demonstrate that autofluorescence-based light-sheet imaging enables reliable visualization of CUBIC-R-cleared mouse embryos, revealing microanatomical features such as nephrons during developmental stages. To support solvent-based clearing protocols, we designed a protective lens cap that shields objectives from solvent erosion and economically converts standard air objectives into high-performance immersion objectives. Using this capped BLX configuration, we achieved cellular-resolution imaging of whole PEGASOS-cleared mouse brains. The system's simplified design and open-source availability promote broader adoption for diverse applications in developmental biology.
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