Positioning of nuclei generated from arrangements of actin filaments. (IMAGE)
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a, Results of the identification and matching of real and generated nuclei by an automatic counting of the whole generated dataset and by manual counting of a subset of images. Stained nuclei refer to those recorded directly using fluorescent microscopy. Generated nuclei are those produced by the neural network using actin filament arrangements. Matched nuclei are those generated at less than 4 μm of its real counterpart. b, Manual (left) and automatic (right) processing of the same image. In manual processing the profile of the nuclei is drawn to calculate the centroid and the nuclei matched by comparison with the real counterpart. On the other hand, the automatic processing automatically identified the nuclei and generated their bounding boxes, matching generated and real nuclei based on the maximization of the overlapping areas of the bounding boxes. c, Several examples of generated nuclei (red) and their corresponding real nuclei (blue). The first three images (green frame) correspond to nuclei generated within the average nuclear radius (4 μm) from their real position. The last image (red frame) corresponds to a mismatch, where the generated nucleus is too far from its real position (See S2 Fig in S1 File for full images; Bars are 20 μm). d, Example of a cell and the relative distance of 4 μm within the cytoplasm. The probability of randomly positioning the nucleus within the cytoplasm can be identified as the ratio of possible matched positions for the centroid (green area) with respect to all possible positions (orange area). Those possible positions of the centroid located at less than the nuclear radius from the edges of the cell (red area) are discarded under the premise that the nucleus cannot be positioned partially outside the cell (See S3 Fig in S1 File for further analysis of probabilities; Bar is 20 μm). e, Distribution of the distances of the generated nuclei respect their real position. 71% of the nuclei are situated at less than 4 μm of their real position. f, Distribution of distances of the generated nuclei considered matched (<4 μm). 40% of the matched nuclei are located at less than 1μm from their real position.
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