2.5. Image segmentation

Images were segmented using Attovision software. Nuclei were counted in DAPI images by using polygon segmentation parameters described in Table 2.1. GM1 aggregates within cells were defined in Texas Red images using polygon segmentation parameters described in Table 2.2. All images were segmented using the same segmentation parameters defined for each filter channel. The segmentation process is depicted in Fig. 1, Fig. 2.

Nuclei segmentation process. After defining dye signal threshold, every image was segmented using the same parameters described in Table 2.1. Nuclei segmentation was used for cell counting.

GM1 aggregates segmentation process. After defining dye signal threshold and applying filters, every image was segmented using the same parameters described in Table 2.2. Pixel area of each aggregate was used to calculate total pixels per image.

Segmentation parameters for nuclei and GM1 aggregates definition.