Focused ion beam-scanning electron microscopy links pathological myelin outfoldings to axonal changes in mice lacking Plp1 or Mag [1151 micrographs in TIFF format]
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Focused ion beam-scanning electron microscopy links pathological myelin outfoldings to axonal changes in mice lacking Plp1 or Mag
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.
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