2.8 Image acquisition and analysis
Image acquisition was performed with a fluorescence laser microscope (Olympus BX51, Germany) using 4X, 10X, 20X objectives, and images were analysed using Imaje J software as previously described. For quantification of amyloid plaques similar and comparable histological areas were selected, focusing on the adjacent positioning of the whole cortical area and the hippocampus. The images were converted to 8-bit grayscale images, thresholder within the linear range and the number of particles (Analyse particle function 10-Infinity), as well as the percentage of area covered by Thioflavin-S (20X objective), was calculated and averaged from two different measurements per animal. Glial fibrillary acidic protein (GFAP) and Iba-1 stained images (10X) were acquired, maintaining constant exposure for all samples across single experiments. The fluorescence intensity of the positive cells was measured in DG, CA1 and CA3 areas of the hippocampus. Quantification was averaged from two to three different measurements.