Enhanced light-sheet illumination by a multi-layered stair-step phase mask

Abstract Light-sheet microscopy is a valuable tool for live biological imaging, offering high speed, high spatial resolution, and low phototoxicity imaging. This study explores a multi-layered stair-step phase mask to further improve illumination of this microscopy. By inserting the mask into the illumination optics, we can modify the Gaussian beam to extend its length by approximately 1.6-fold, which increases the field of view in light-sheet microscopy while keeping resolution reduction small. We demonstrate this improvement through in vivo imaging of medaka fish embryos, highlighting the mask’s potential to enhance the visualization of multicellular organisms at the cellular level.