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Chemotaxing Dictyostelium discoideum Cells

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Differentiated Dictyostelium cells were spotted on a chamber slide and allowed to aggregate. After fixing, staining and resin embedding, pairs of trailing cells in close proximity to each other were located and imaged by focused ion beam scanning electron microscopy (FIB-SEM). The raw image stack was aligned, binned and contrast-inverted to yield an image volume with 15 nm isotropic (cubic) pixel sampling. These image volumes were examined for the presence of exosomes and were used for segmentation and 3-D reconstruction of these structures and the Dictyostelium cells themselves.