Rapid adaptive optical recovery of optimal resolution over large volumes

Adaptive optics microscopy using a de-scanned, laser-induced guide star and direct wavefront sensing allows high numerical aperture diffraction-limited imaging of fine dynamic structures deep in the intact living zebrafish brain. Using a descanned, laser-induced guide star and direct wavefront sensing, we demonstrate adaptive correction of complex optical aberrations at high numerical aperture (NA) and a 14-ms update rate. This correction permits us to compensate for the rapid spatial variation in aberration often encountered in biological specimens and to recover diffraction-limited imaging over large volumes (>240 mm per side). We applied this to image fine neuronal processes and subcellular dynamics within the zebrafish brain.