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Selective plane illumination microscopy (SPIM) offers an alternative way of optically sectioning the sample in fluorescence microscopy. By illuminating the sample with a sheet of light, a sectioning effect can be obtained that is similar to that in confocal microscopy. However, in combination with widefield detection, such an approach has several advantages over confocal scanning microscopy. SPIM offers reduced fluorophore1 bleaching, fast, highly efficient image recording, and high depth penetration, especially when multiple views are combined. SPIM performs especially well in large samples such as fish or fly embryos, which can be observed live for several days. The principle is universal and can also be applied to micrometre-sized samples.

Studying live embryos in 4D: Selective plane illumination microscopy: a new technique for high-resolution four-dimensional imaging