Beam shaping in light-sheet microscopy: an experimental analysis

Thanks to its unique optical sectioning capability, light-sheet fluorescence microscopy has proven
to be a powerful technique for volumetric imaging of entire model organisms with high spatial and
temporal resolution. For light sheet generation with scanned laser beams, holographic beam
shaping offers precise control over the optical fields exciting the fluorescence. Various illumination
schemes have been proposed, aiming for best image quality with regard to axial resolution, optical
sectioning, illumination homogeneity and photobleaching while at the same time retaining a large
field of view. Here, we have engineered and characterized a variety of beams and analyzed their
imaging performance by using phantom samples and zebrafish embryos. These data may assist
researchers to select the light sheet best suited to the imaging application at hand.