3D high- and super-resolution imaging using single-objective SPIM

Light-sheet microscopy using a single objective and micromirrors to orient the light sheet enables high- and super-resolution imaging of cells and embryos on a standard microscope. Single-objective selective-plane illumination microscopy (soSPIM) is achieved with micromirrored cavities combined with a laser beam–steering unit installed on a standard inverted microscope. The illumination and detection are done through the same objective. soSPIM can be used with standard sample preparations and features high background rejection and efficient photon collection, allowing for 3D single-molecule-based super-resolution imaging of whole cells or cell aggregates. Using larger mirrors enabled us to broaden the capabilities of our system to image Drosophila embryos.