Histological Analysis of Brain Samples

For histological studies, the brains isolated from experimental animals were fixed in a 10% formalin solution for 24 h at room temperature. For the next 24 h, the samples were placed in a 15% sucrose solution followed by incubation in a 30% sucrose solution for 24-48 h. After that, the brain was placed on a platform of freezing sliding cryostat Leica CM1520 (Leica, Germany) and gradually filling with cryogel (Leica, Germany) at 30° C. Next, the sample was cut into 10 μm thin coronal sections. Every fifth section was mounted on a glass slide and dried in the air for 24 h. The sections were then stained by the hematoxylin-eosin method [83 (link)]. Then, the sections were dehydrated in alcohols of upward concentration, purified in xylenes and embedded in a mounting medium (Consul-Mount, USA). The samples were examined using a Zeiss Primo Star light microscope (Germany) with integrated an Axio CamMRc camera (Zeiss, Germany).