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Multiphoton microscopy applied in bone tissue is susceptible to optical aberrations caused by heterogeneity in refractive index. Optical clearing can be applied to alleviate some of these aberrations, but it is invasive and causes deviations from normal tissue biology. We recover diffraction limited imaging by means of a high spatial frequency digital micromirror device (DMD), and binary wavefront modulation. A genetic algorithm optimizes the DMD pattern by evaluating the intensity of the Second Harmonic Generation point spread function measured in the bone sample. We present a five-fold GFP intensity improvement, and a 29% spatial resolution increase within an ex vivo mouse sample.
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Adrian Liversage, Kayvan Forouhesh Tehrani, Tianyi Zheng, Peter Kner, Luke Mortensen, "Binary wavefront manipulation using an epi-detected SHG guidestar," Proc. SPIE 12851, Adaptive Optics and Wavefront Control for Biological Systems X, 1285107 (12 March 2024); https://doi.org/10.1117/12.3003515