Image segmentation consists of partitioning the image into different objects of interest. For a biological image, the segmentation step is important to understand the biological process. However, it is a challenging task due to the presence of different dimensions for cells, intensity inhomogeneity, and clustered cells. The marker-controlled watershed (MCW) is proposed for segmentation, outperforming the classical watershed. Besides, the choice of markers for this algorithm is important and impacts the results. For this work, two foreground markers are proposed: kernels, constructed with the software Fiji and Obj.MPP markers, constructed with the framework Obj.MPP. The new proposed algorithms are compared to the basic MCW. Furthermore, we prove that Obj.MPP markers are better than kernels. Indeed, the Obj.MPP framework takes into account cell properties such as shape, radiometry, and local contrast. Segmentation results, using new markers and illustrated on real Drosophila dataset, confirm the good performance quality in terms of quantitative and qualitative evaluation.