Anti aβ42

Coronal sections (40 μm thick) were incubated overnight at 4°C with anti-Aβ₄₂ (Dr. Vitaly Vasilevko and Dr. David H. Cribbs, University of California, Irvine), anti-GFAP (Millipore, Billerica, MA) or anti-CD45 (AbD Serotec, Raleigh, NC) with 5% normal serum in Tris-buffered solution. The sections were subsequently treated with the appropriate biotinylated secondary antibody; then processed using the Vectastain Elite ABC reagent and 3,3′-diaminobenzidine (Vector Laboratories, Burlingame, CA), according to the manufacturer’s instructions. Sections from vehicle and ATL treated mice were processed under the same conditions.
The immunostaining was assessed over a region representing a distance of approximately 160 μm obtained between 1.34 and 2.54 mm posterior to the bregma. Images of stained hippocampus, entorhinal cortex, subiculum, and amygdala were acquired using an Axiocam digital camera and AxioVision software version 4.6 connected to an Axioskop 50 microscope (Carl Zeiss MicroImaging, Thornwood, NY). Settings for image acquisition were identical for vehicle and ATL treated tissues.
Staining analyses were calculated as the percentage of labeled area captured (positive pixels)/the full area captured (total pixels) using ImageJ complying with strict standards [22 (link)].