Brain hippocampal and cortical sections were pre-treated for 1 h using the 2100 antigen retriever (Aptum biologics Ltd, Southampton, UK) followed by 90%formic acid then 0.1%triton-X treatment for 5 min and 1 min, respectively. Sections were then blocked with 0.3%H2O2 for 15 min to inactivate endogenous peroxidases and with Protein Block Serum-Free (Agilent, Santa Clara, CA, USA) for 15 min before adding the primary antibodies. Sections were then stained with the following primary antibodies: 4G8 anti-Aβp (1:500; Bio legend, San Diego, CA, USA) and AT8 anti-p-Tau (Ser202, Thr205) antibody (1:500; Thermo-fisher Scientific, MA, USA) respectively for 1 h at room temperature (RT). After washing three times with Tris buffered saline/0.05%Tween (TBST), sections were stained with secondary antibodies in TBS: HRP-conjugated anti-mouse IgG (1:500; Sigma-Aldrich) or anti-rabbit IgG (1:500; Sigma-Aldrich) respectively for 1 h at RT. Sections were again washed three times with TBST before addition of 3,3’-Diaminobenzidine (DAB) substrate chromogen system and incubated for 5–10 min. Slides were then counterstained with hematoxylin for 1 min. After mounting, slides were finally imaged using the Olympus CX 43 light microscope (Shinjuku, Tokyo, Japan) [20 (link)].