Histopathological study was performed as a previously described routine method [12 (link)]. Briefly, brain tissues were fixed in 4% neutral buffered
paraformaldehyde and washed with tap water for overnight. They were dehydrated in a graded series of ethyl alcohol (70 to 100%), cleaned with xylene, and embedded in paraffin with routine
protocol. Paraffin blocks were sectioned at a thickness of 4 µm using a rotatory microtome (Leica, Wetzlar, Germany). Sections were mounted on slide glass and dried on slide
warmer. They were immersed in xylene, rehydrated in a graded series of ethyl alcohol (100 to 70%), and washed with water. Sections were stained with Harris hematoxylin solution (Sigma
Aldrich) and eosin Y solution (Sigma Aldrich). After staining, sections were dehydrated with a graded ethyl alcohol series, cleaned in xylene, and subsequently mounted with mounting media
(Thermo Fisher Scientific, Waltham, MA, U.S.A.). Mounted sections were observed with Olympus microscope (Olympus, Tokyo, Japan).
paraformaldehyde and washed with tap water for overnight. They were dehydrated in a graded series of ethyl alcohol (70 to 100%), cleaned with xylene, and embedded in paraffin with routine
protocol. Paraffin blocks were sectioned at a thickness of 4 µm using a rotatory microtome (Leica, Wetzlar, Germany). Sections were mounted on slide glass and dried on slide
warmer. They were immersed in xylene, rehydrated in a graded series of ethyl alcohol (100 to 70%), and washed with water. Sections were stained with Harris hematoxylin solution (Sigma
Aldrich) and eosin Y solution (Sigma Aldrich). After staining, sections were dehydrated with a graded ethyl alcohol series, cleaned in xylene, and subsequently mounted with mounting media
(Thermo Fisher Scientific, Waltham, MA, U.S.A.). Mounted sections were observed with Olympus microscope (Olympus, Tokyo, Japan).
