Bx 51 binocular research microscope

Animals were perfused with 0.9% dextrose saline followed by 4% paraformaldehyde (PFA). Brain tissues were excised and fixed in 4% w/v paraformaldehyde. Transverse sections (5–6 μm thick) were obtained from the PFC and CA3 region of the hippocampus using a microtome (Leica, Germany), and the sections were fixed on glass slides. Hematoxylin and eosin (H&E) staining was carried out to demonstrate the general histological profiles of the brain regions according to the method described (
Eltony & Elgayar, 2014). Images were acquired using an Optronics digital camera connected to a computer interface (MagnaFire) and an Olympus BX-51 binocular research microscope. The general structure of the pyramidal cells, periglomerular, and granule cells was characterized using inter-reader variability. Viable neuronal cells were defined as round-shaped, cytoplasmic membrane-intact cells without any nuclear condensation or distorted aspect. The number of viable neurons was determined using ImageJ software and counted as a ratio of viable neuronal cell counts to the square area of the circular view in a section.

The images were acquired using an Optronics Digital Camera connected to a computer interface (MagnaFire) and an Olympus BX-51 Binocular research microscope. The general structure of the pyramidal cells peri-glomerular and granule cells were characterized using inter-reader variability. The cells were counted using Image J at X400 or X250. Immunopositive cells (p53 and Bax) were counted at different microscopic fields (n = 7) for n = 5 sections for all groups using the method of Going, (1994).^{31 (link)} Data obtained was analyzed using ANOVA and Bon Ferroni Post Hoc test with significance set at P<0.05 [Graph Pad Prism (Version 6.0)]. A p-value of less than 0.05 was considered statistically significant, thus, P<0.05 (*), P<0.01 (**) and P<0.001 (***).