REL 2022-09-15 2022-12-13 2022-12-13 0000-0002-4814-7517 Anna Maria Steyer European Molecular Biology Laboratory Meyerhofstrasse 1 Heidelberg Baden-Württemberg Germany 69117 0000-0002-7710-5738 Hauke Werner Department of Neurogenetics, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany Hermann Rein Str. 3 Göttingen Lower Saxony Germany 37075 Steyer AM Möbius W 17.8 10.6019/EMPIAR-11215 FIB-SEM tissue Steyer AM Buscham TJ Lorenz C Hümmert S Eichel-Vogel MA Schadt LC Edgar JM Köster S Möbius W Nave KA Werner HB Glia Glia 2022 English 10.1002/glia.24290 36354016 EMPIAR-11214 EMPIAR-11220 EMPIAR-11219 EMPIAR-11237 EMPIAR-11238 EMPIAR-11239 EMPIAR-11240 EMPIAR-11216 FIB-SEM of mouse optic nerve of Plp knock-out (Plp-/Y-4017) /data micrographs - single frame TIFF TIFF 1151 1 UNSIGNED BYTE 3984 50.0 4176 50.0

Optic nerve of mice harboring null-alleles of the Plp/Plp1 (Plp-/Y) sarificed 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 (600 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 2), local contrast enhancement (127, 55, 1.5) and sharpened with Fiji/ImageJ.