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In keeping with Vogt's method, each brain is cut into four or five chunks, which are encased in paraffin wax.
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Each rat brain was cut into six sections and mouse brain into five sections.
The brain was cut into 7 µm sections which were stained with hematoxylin and eosin (H&E stain).
For tissue analyses, the brain was cut into 10 µm coronal sections on a cryostat (Leica Microsystems).
The unfixed brain was cut into frontal slabs of approximately 10 mm thickness.
Coronal sections of the entire brain were cut into 40-μm sections using a cryostat.
The brain was cut into slices (30 μm thick) using a cryostat and the sections were placed on glass slides.
Coronal sections of the brain were cut into 40 50 μM thickness using a vibrating microtome (VT1000S, Leica, Milton Keynes, UK).
The remaining brain was cut into 350 μm thick horizontal slices using a vibratome (Leica VT 1000S, Bensheim, Germany).
The anterior part of the brain was cut into coronal sections of 40 μm and stored for ChAT immunohistochemistry.
Frontal lobes and cerebellum were dissected of the hemispheres and the remaining brain was cut into 350-μm-thick horizontal slices with a vibratome (VT1000S; Leica, Bensheim, Germany).
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