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Optic nerve of wild type (littermates) mice sacrificed by cervical dislocation at postnatal day 75 (P75). The optic nerves were dissected and immersion fixed and prepared using the reduced osmium tetroxide – thiocarbohydrazide - osmium (OTO) method as previously described in detail (Weil et al. 2018; Steyer et al. 2020) upon implementing modifications to the original protocol first introduced by (Deerinck et al. 2010). The micrographs are obtained by FIB-SEM (Zeiss Crossbeam 540): SEM: 1.5 kV (1000 pA), ESB grid 450V, pixel size x/y 5 nm, continuous mill-and-acquire mode; FIB: 1.5 nA , z-step 25 nm. Postprocessing: alignment with Fiji/ImageJ SIFT, cropped, inverted and blurred (Gaussian blur, sigma 1), local contrast enhancement (127, 256, 1.5).

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