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Hi, Im Anne Carpenter from the Broad Institute of Harvard and MIT, and I co-lead the CellProfiler project. Sometimes you can answer your biological question of interest by measuring properties of the entire image, such as the total amount of fluorescence, or the texture or smoothness of staining across the entire field of view. But thats fairly rare. Usually, youre going to need to measure individual biological entities with image within the image, and those entities can be things like nuclei, cells, synapses, vesicles. They could be regions of organisms or tissue samples, or colonies growing on a plate. Segmentation is the process of identifying individual cells or structures within the image. And for most bioimage analysis workflows, segmentation is a key step. Segmentation gets its name from segmenting or dividing the pixels in the image into regions of interest, otherwise called ROIs, such as individual cells, as shown here. In some software, this step may be called segmenta