Image analysis of fluorescence signals. A). Automated quantitative image analysis through the tile-sampling classifier. Non-overlapping square tile were placed on DAPI-counterstain image in order to maximize the nuclear material covered. Each tile may contain a single nucleus or portions of one or more nuclei. The software effectuated the spot count in each tile. B). Quantitative image analysis through the automated nuclei-sampling classifier. The classifier automatically recognized individual nuclei in counterstain image (same image as in Figure 1A). Nuclei that were automatically recognized and considered in the analysis showed green outlines, while nuclei that were automatically recognized but not considered in the analysis had a white outline. The software effectuated spot count in each nucleus. C). After completion of the automated image analysis, the user could improve the selection of automatically selected nuclei via interaction with the software. In this particular case, some nuclei that were not yet considered during the automatic segmentation (Figure 1B) were selected by the user, some automatically selected nuclei were deleted and some overlapping nuclei were divided (yellow line between two overlapping nuclei).
Furrer et al. Diagnostic Pathology 2013 8:17 doi:10.1186/1746-1596-8-17