Curved axially scanned light-sheet microscopy

Abstract Light-sheet fluorescence microscopy enables high-throughput multidimensional imaging. However, attempts to further improve throughput by incorporating commercially available or custom high space-bandwidth product objectives have been limited by objective field curvature, which violates the co-planar overlap required for light-sheet imaging and ultimately reduces the usable field of view. Inspired by machine-vision strategies that curve the image plane to match the field curvature, we introduce curved axially scanned light-sheet microscopy, which adapts the light-sheet excitation to the detection objective’s field curvature via synchronized control of the remote refocus scan and motorized mirror. Using our technique, we increase the usable field of view along the light-sheet propagation axis from ∼2.5 mm to ∼6.3 mm for a commercial high-SBP detection objective with significant field curvature, while maintaining sub-cellular resolution.