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We propose an adaptive optics light-sheet fluorescence microscope (AO-LSFM) for closed-loop aberrations’ correction at the emission path, providing intrinsic instrumental simplicity and high accuracy when compared to previously reported schemes. The approach is based on direct wavefront sensing, i.e., not on time-consuming iterative algorithms, and does not require the use of any guide star, thus reducing instrumental complexity and/or sample preparation constraints. The design is based on a modified Shack–Hartmann wavefront sensor providing compatibility with extended sources such as images from optical sectioning microscopes. We report an AO-LSFM setup based on such sensors, including characterization of the sensor performance, and demonstrate for the first time to the best of our knowledge a significant contrast improvement on neuronal structures of the ex vivo adult drosophila brain in depth.

We report, for the first time to our knowledge, experimental demonstration of wave-front analysis via the Hartmann technique in the extreme ultraviolet range. The reference wave front needed to calibrate the sensor was generated by spatially filtering a focused undulator beam with 1.7- and 0.6-μm -diameter pinholes. To fully characterize the sensor, accuracy and sensitivity measurements were performed. The incident beam’s wavelength was varied from 7 to 25 nm. Measurements of accuracy better than λ_EUV/120 (0.11 nm) were obtained at λ_EUV=13.4 nm . The aberrations introduced by an additional thin mirror, as well as wave front of the spatially unfiltered incident beam, were also measured.