Ten brains (five females with age range of 59–86 years, and five males with age range of 30–75 years, mean age of 65, 8 years) were obtained from the body donor program of the Department of Anatomy at the University Hospital Düsseldorf of the Heinrich-Heine-University in accordance with legal requirements with no indications of neurologic or psychiatric diseases in clinical records. The postmortem delay did not exceed 24–36 h (Table 1 ). The brains were fixed in 4% buffered formalin (pH 7.4) or Bodian’s fixative for at least 6 months. All brains underwent magnetic resonance imaging on a Siemens 1.5 Tesla scanner (Erlangen, Germany) using a T1-weighted 3D FLASH sequence (flip angle 40°, repetition time TR 40 ms, echo time TE 5 ms). Obtained images were used as an undistorted spatial reference for the 3D-reconstruction of the histological sections as previously described (Bludau et al., 2014 (link)).
Brains were embedded in paraffin and serially sectioned in the coronal plane on a large-scale microtome (thickness of 20 μm). Every 15th section (corresponding to a distance of 300 μm) was mounted on a glass slide covered with gelatin, stained for cell bodies using a silver staining technique (Merker, 1983 (link)), and digitized on a flatbed scanner (resolution of 1,200 dpi). At least every 60th section was analyzed (distance between them of 1.200 μm).
Brains were embedded in paraffin and serially sectioned in the coronal plane on a large-scale microtome (thickness of 20 μm). Every 15th section (corresponding to a distance of 300 μm) was mounted on a glass slide covered with gelatin, stained for cell bodies using a silver staining technique (Merker, 1983 (link)), and digitized on a flatbed scanner (resolution of 1,200 dpi). At least every 60th section was analyzed (distance between them of 1.200 μm).
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