Experimental light-sheet microscopy for the comparison of different methods of structured illumination

Light-sheet microscopy is an established method to nondestructively visualize small biological structures in three dimensions. The specimen needs to be chemically treated (e.g. decalcified, cleared to transparency) and dyed with a fluorescence dye. The microscope produces a spatially limited illumination field (usually a light sheet), which only excites the fluorescence dye in the focus plane of the detection microscope, which is mounted perpendicularly to the illumination path. Therefore less light is emitted by the other parts of the specimen and the image quality increases. Commercial systems as well as projects like OpenSPIM or the sTSLIM in our partner`s lab in Minneapolis are highly specialized and complicated systems optimized for the needs of biological and medical research. Their setups do not, respectively do not easily allow to experiment with various methods of producing structured illumination. We present a basic setup of a light-sheet microscope which allows us to thoroughly simulate and characterize the influence of different illumination methods on the imaging quality.