*4.3. Otsu Binarization*

After grayscale conversion, segmentation is made with automatic threshold to separate cells, as shown in Figure 7. To improve image quality and correct any defects due to Otsu's binarization, such as holes in cell nuclei, the process benefits from the use of aperture in combination with dilation.

**Figure 7.** Output images of Otsu's binarization (**a**) and morphological operations (**b**).
