BENCHTOP LIGHT-SHEET MICROSCOPE FOR MESOSCOPIC IMAGING OF CLEARED TISSUE

Light-sheet microscopy enables rapid volumetric imaging with reduced photodamage and labour cost compared to confocal approaches and conventional sectioning. However, adoption of light-sheet microscopy remains limited by the cost and footprint of commercial systems and previous open-source designs.
We developed a compact benchtop light-sheet microscope that fits within a 45 cm × 30 cm breadboard. The optical design integrates Powell lens–based light-sheet generation within a folded path, supported by passive vibration dampening for benchtop imaging. Control and acquisition were implemented using a single-board computer, stepper motor controllers, and an embedded machine vision camera, providing automated scanning and synchronised image capture. The modular assembly design reduces alignment complexity, while remaining adaptable to user-specific requirements. Assembly tools can be 3D‑printed, with designs deposited in community repositories ensuring reproducibility.
The system achieved a cellular resolution with resolving power of 3.91µm, measured by imaging of full-width-half-maximum of standard 1µm fluorescent beads. Whole-brain imaging of cleared mouse tissue injected with viral tracers was completed in under 8 minutes. Injection sites were localised and mapped to the Allen Mouse Brain Atlas, demonstrating the system’s ability to support rapid histological imaging for most neuroanatomical analyses.
This benchtop microscope provides an accessible open-source solution for mesoscopic imaging, bridging the gap between local laboratory setups and central facility systems. Its compact design, rapid acquisition, and reproducibility, expand experimental capacity in settings where space and resources are limited, supporting rapid, low photodamage, large-scale histological applications.