Automatic white blood cell segmentation using stepwise merging rules and gradient vector flow snake

► Propose two different schemes for segmenting the nuclei and cytoplasm of WBCs. ► For nuclei segmentation, a probability map is created using a probability density function estimated from samples of WBC's nuclei and sub-images cropped to include a nucleus based on the fact that nuclei have a salient color against the background and red blood cells. Mean-shift clustering is then performed for region segmentation, and a stepwise merging scheme applied to merge particle clusters with a nucleus. ► For cytoplasm segmentation, morphological opening is applied to a green image to boost the intensity of the granules and canny edges detected within the sub-image. The boundary edges and noise edges are then removed using removal rules, while a GVF snake is forced to deform to the cytoplasm boundary edges. ► The proposed method is demonstrated to improve the segmentation performance when compared to another method. ► Experimental results confirm that the proposed method can efficiently segment the nucleus and cytoplasm of WBCs, regardless of the WBC category. This study aims at proposing a new stained WBC (white blood cell) image segmentation method using stepwise merging rules based on mean-shift clustering and boundary removal rules with a GVF (gradient vector flow) snake. This paper proposes two different schemes for segmenting the nuclei and cytoplasm of WBCs, respectively. For nuclei segmentation, a probability map is created using a probability density function estimated from samples of WBC's nuclei and sub-images cropped to include a nucleus based on the fact that nuclei have a salient color against the background and red blood cells. Mean-shift clustering is then performed for region segmentation, and a stepwise merging scheme applied to merge particle clusters with a nucleus. Meanwhile, for cytoplasm segmentation, morphological opening is applied to a green image to boost the intensity of the granules and canny edges detected within the sub-image. The boundary edges and noise edges are then removed using removal rules, while a GVF snake is forced to deform to the cytoplasm boundary edges. When evaluated using five different types of stained WBC, the proposed algorithm produced accurate segmentation results for most WBC types.