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Joana Silva, Isabel F. Amaral, Pedro Quelhas (INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Portugal)The main goal of this work is to improve the visualization and quantitative analysis of radial cell outgrowth from neurospheres embedded or plated on three-dimensional hydrogels, to assess the effect of soluble factors/ECM proteins/immobilized ligands on neural progenitor cell migration. Such outgrowth quantification is traditionally quantified based on a 2-D maximal projection of the confocal microscopy stack, both possibly collapsing diverse outgrowth into one and ignoring different vertical outgrowth angles. In our work fluorescently labelled nuclei and cytoplasm were imaged in a confocal laser scanning microscope leading to a 3-D stack of images of the neurospheres, where nuclei clustering marks the neurosphere and cytoplasmic processes with few nuclei are considered to be outgrowth. Using 3D segmentation and image morphology the respective volumes were selected, enabling a 3D visualization and the 3D computation of the maximal outgrowth distance and outgrowth volume.Acknowledgements: Portuguese FCT project (PTDC/SAU-BMA/118869/2010),FEDER through COMPETE (Pest-C/SAU/LA0002/2013).