Fig. 1

(a) 3D model of the complete setup. (b) Slow scan mode (Left): The unit is mounted on the motorized stage of the microscope that moves the objective, enabling slow z-scans through the sample while keeping the light sheet in the imaging focal plane. Fast scan mode (Right): Synchronized actuation of the piezo positioners, which hold the objective and fiber, enables fast, high-precision z-scans. (c) Step-by-step construction plan of the light-sheet unit. (d) Cross-sectional view of the light-sheet unit, illustrating the galvanometric mirror that scans the laser beam in the sample plane to generate the digitally scanned light sheet. (e) Alignment procedure via mechanical manipulation of the entire light-sheet unit. Left: The unit (LU) is moved vertically using a mechanical z-stage (indicated by the yellow arrow) to align the light sheet with the imaging objective’s focal plane. Right: The LU is moved horizontally using a mechanical x-stage (indicated by the yellow arrow) to position the light-sheet waist at the center of the imaging objective’s field of view. 3D design software: Autodesk Fusion360 v2.0.