Multistage Morphological Segmentation of Brightfield and Fluorescent Microscopy Images

This paper describes the multistage morphological segmentation method (MSMA) for microscopic cell images. The proposed method enables to study the cell behavior using a sequence of two types of microscopic images: bright field images and/or fluorescent images. The proposed method is based on two types of information: the cell texture coming from the bright field images and intensity of light emission, done by fluorescent markers. The method is dedicated to the image sequences segmentation and is based on mathematical morphology methods supported by other image processing techniques. The method allows detecting cells in image independently from a degree of their flattening and from presenting structures which produce the texture. It makes use of some synergic information from the fluorescent light emission image as the support information. The MSMA method has been applied to images acquired during the experiments on neural stem cells as well as to artificial images. In order to validate the method, two types of errors have been considered: the error of cell area detection and the error of the cells’ position using artificial images as the “gold standard”.