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.