Hello Nisreen,

Please go through these points and the attached file regarding this,

A preprocessing step is used to remove the background and uniform the light intensity conditions, then the segmentation process takes place with a two-dimensional version of the Enhanced Interaction Model. The resulting binary image is then enhanced through a cascade of morphological operators. Different colonies are then processed through a Watershed transform which returns as output the segmentation for each colony in the image.

Please go through this