Flowchart of the overall procedure. The first step consists in training the program by learning sample nuclei. The user clicks on the approximate position of the nuclei in the image. Using an iterative thresholding algorithm, nuclei are automatically extracted around each clicked position. The user has to validate the segmented nuclei that will serve as training samples for the classifier. The second step is completely automatic and will segment, as well as classify, objects in the entire dataset.