ABSTRACT
Light sheet microscopy enables high-speed, three-dimensional imaging for extended periods of time, and when this is linked with biological techniques, such as the use of fluorescent proteins, dynamic processes can be studied with both high temporal and spatial resolution. The original method, using a sheet of light rather than the more conventional patch or single spot, was known as “ultra-microscopy”. This chapter looks at the history of the development before explaining the basic optical principles together with the design and application of a practical system and the basic configurations. The fundamental optical concepts of the technique are simple; requiring just basic optics and a camera, therefore high-performing instruments can be built easily in a user’s laboratory for a reasonable price. There are numerous variations of selective plane illumination microscopy (SPIM) systems, but as with most forms of optical microscopy there is one decision that does need to be made early on, that of conventional or inverted SPIM.
