Fig. 1.

Procedures of the histogram-fitting-based analysis for confocal microscope images. A representative microscope image of a male 5xFAD mouse is utilized for demonstration. The whole slice stained with Iba1 (A), CD68 (B), and DAPI (C) is displayed. (D) and (E) represent the histograms of Iba1- and CD68-stained signals; the x-axis represents pixel signal intensity, while the y-axis represents the pixel count. (F) Mask corresponding to the signal intensity of 0; (G) 1st Gaussian fitting for extracting background signals (µ_Histo1 = 17); (H) Mask corresponding to signal intensities ranging from 0 to 17; (I) Zoom-in vision of H overlaid on the Iba1-stained image; (J) 2nd Gaussian fitting for extracting signals of interest (µ_Histo2 = 49); (K) Mask corresponding to signal intensities larger than 49; (L) Zoom-in vision of K overlaid on the Iba1-stained image.