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To address the lack of systematic documentation for customizing optical hardware to diverse specimen requirements, the authors iteratively re-engineered the open-source OpenSPIM platform into an expanded, modular light-sheet microscope. The design integrates multi-wavelength laser illumination, configurable emission filtration, and specialized sample-handling workflows. Culminating in FlowSPIM, the platform successfully combines continuous flow-based fluidic incubation with volumetric light-sheet imaging for extended 3D live-tissue culture observation.
Open-source light sheet microscopy can break free of rigid commercial constraints through modular iterative design, yielding a flow-incubation imaging system capable of continuous 3D tissue culture observation.
Light sheet microscopy is ideal for the 3D imaging of large biological specimens, yet systematic documentation on adapting microscope designs to diverse experimental requirements remains limited. We present the iterative process used to design and build a modular light-sheet microscope and adapt it to different sample types and experimental conditions. Starting from a stripped-down OpenSPIM platform, we gradually make our microscope more versatile by implementing new hardware, such as additional laser channels and optical filters, and new protocols for sample handling. In its final iteration, FlowSPIM is a new flow-based incubation technique for 3D tissue culture.