Combined Detection and Segmentation of Overlapping Erythrocytes in Microscopy Images Using Morphological Image Processing

Abstract

Segmentation of clusters of erythrocytes into their constituent single cells is a procedure needed in various biomedical applications related to microscopy images. This task is part of the general problem of splitting clumps of objects in images which continues being an open research topic in the Image Processing field. This work presents a unified morphological method to detect and segment clusters of erythrocytes in microscopy images, and proposes two main contributions. The first one is to formulate and evaluate a method to detect clusters as connected components in binary images, obtained from a previous coarse segmentation, which is not capable of further dividing a cluster into its constituent cells. Secondly, to propose the best alternative to split the clusters into their constituent individual cells after evaluating three algorithms based in the combination of the transforms: H-maxima, weighted external distance and marker-controlled watershed. Evaluation of the proposed cluster detection methods was made in terms of standard measures of effectiveness. Segmentation accuracy was evaluated comparing the segmented objects obtained to a manually segmented ground truth, by means of the Jaccard index. Then the Friedman test allowed validating the advantages of the proposed method in comparison to the other alternatives studied here.