OmniSphero comprises a classical watershed algorithm to identify nuclei and to obtain information like position of the centroid of the nucleus on the image, nucleus area and intensity. The coordinates of the nuclei centroids are later used to associate a certain nucleus with a cell type (e.g. neurons). Additionally OmniSphero is able to import coordinates of nuclei from other software and to use for analyzing other endpoints such as neuronal identification or radial migration distance. In the case of neuronal identification this allows OmniSphero to perform a one by one comparison of identified nuclei positions associated to neurons between its own algorithms and algorithms of other software.
Figure 3: The left image shows an image from the nucleus channel (Hoechst stained nuclei, represented in green). The image is thresholded converted to a binary image and consequently a watershed algorithm is used to segment overlapping nuclei. Finally coordinates of centroids of identified nuclei are safed and can be displayed as a blue points cloud.