Thin-sheet laser imaging microscopy

We present a general summary of light sheet based microscopy for non-destructive optical sectioning of organisms and thick tissues. Optical sections are recorded using a thin sheet of light to induce a plane of fluorescence in transparent or fixed and cleared tissues. We explain the basic building units and compare our thin sheet laser imaging microscope (TSLIM) to similar microscope systems. High resolution across the full width of a large specimen is achieved by moving the specimen through the thinnest region of the hyperbolically shaped light sheet, stitching together pixel columns with optimal resolution. We also show how to reduce absorption and scattering artefacts and explain optimizations to the optical system which help to produce thinner light sheets than achieved with cylindrical lenses alone.