Compact, high-speed multi-directional selective plane illumination microscopy

Abstract

We present an elegant scheme for providing multi-directional illumination in selective plane illumination microscopy (SPIM). Light sheets can be delivered from one of two opposed directions at a time and pivoted about their center for efficient stripe artifact suppression using only a single galvanometric scanning mirror to perform both functions. The scheme results in a much smaller instrument footprint and allows multi-directional illumination with reduced expense compared with comparable schemes. Switching between the illumination paths is near instantaneous and the whole-plane illumination scheme of SPIM maintains the lowest rates of photodamage, which is often sacrificed by other recently reported destriping strategies. The ease of synchronization allows this scheme to be used at higher speeds than resonant mirrors typically used in this regard. We provide validation of this approach in the dynamic environment of the zebrafish beating heart, where imaging at up to 800 frames per second is demonstrated alongside efficient suppression of artifacts.