Beam shaping in light-sheet microscopy: an experimental analysis

Abstract

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.

Keywords

• light-sheet microscopy
• beam shaping
• fluorescence microscopy