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Abstract
Fluorescence microscopy provides insights into cellular structure and function, but the undesirable bending of light from the sample or imperfect optics (“optical aberrations”) often degrade imaging resolution or signal. Adaptive Optics (AO) techniques that sense and subsequently correct the aberrations can restore diffraction-limited imaging, but most implementations remain technically complex and expensive. Wavefront sensing, the key step in AO, remains particularly challenging due to slow speed or additional illumination dose imparted to the sample. Previously we showed that phase diversity (PD), a method for wavefront sensing originally developed for use in astronomy, can rapidly sense and correct aberrations in a widefield fluorescence microscope. Here we extend this approach to light sheet microscopy, showing that the PD-based AO of sample-induced aberrations in live Caenorhabditis elegans embryos restores image contrast and resolution, enabling subcellular imaging throughout the imaging volume. In addition to these preliminary results, the further improvement and extension of this technology to additional samples and microscopes will be discussed.
