Cell counting plays a vital role in biomedical researches. However, manual cell counting is time-consuming, laborious, and low efficiency and has a high counting error rate problem. An automatic counting approach for Hela cells of phase-contrast microscopic images is proposed based on the combination of Otsu and watershed segmentation methods to solve the mentioned issues. Firstly, image preprocessing is performed. Secondly, the Otsu method was used to obtain an automatic global optimal threshold for segmentation to achieve batch counting of images. Thirdly, the marker watershed was performed to separate adherent cells and to avoid oversegmentation simultaneously. Finally, cells in phase-contrast microscopic images were counted by detecting the numbers of connected domains in the binary image.

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