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The spinal cord and brain were embedded in paraffin and cut into serial 6-µm thick coronal slides.
Coronal sections (4 μm) through the brain were embedded in paraffin and stained with hematoxylin and eosin (H&E) for microscopic evaluation of neuronal damage.
Rat brains were fixed by transcardial perfusion with saline, followed by perfusion and immersion in 4% paraformaldehyde, and the brain were embedded in paraffin and cut into 5 μm sections.
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"The entire brain is embedded in a dense meshwork of serotonin-releasing fibers, but only a small part of the brain is directly involved in the feelings that we can consciously access be it sadness or joy," Janusonis told VICE.
The left hemisphere of the brain was embedded in tissue freezing medium (Triangle Biomedical Sciences, Durham, NC) and rapidly frozen in a 2-methylbutane/dry-ice bath.
Brains from WT and mutant animals were harvested and the unfixed brain was embedded in OCT, frozen on dry ice, sectioned at 15 µm on a cryostat (Leica), mounted on glass slides and stored at −80°C until used.
Frozen brain was embedded in cryomatrix compound (Thermo Fisher Scientific, Waltham, MA) prior to sectioning.
For sectioning, the brain was embedded in OCT compound (Sakura, Torrance, CA, USA) and sectioned in the sagittal plane.
Following fixation the brain was embedded in optimal cutting compound (OCT) and 8μm coronal sections were made using a cryostat.
Anaesthetized male mice at post-natal day 56 were fixed by perfusion with tissue fixative (GenoStaff, Tokyo, Japan) and the whole brain was embedded in paraffin.
Then the brain was embedded in gelatin/albumin, postfixed in 4% buffered formaldehyde for 17 h at 4 °C, and sectioned at 40 μm with a vibrating blade microtome (Leica VT 1000S; Leica, Nussloch, Germany).
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