The high values of adhesion, the smooth surface topography and the decreased number of protrusions observed in the SW620 cells suggested an altered actin organization, which has been then analysed by confocal imaging (Fig. 1.3). In Fig. 1.3 representative confocal micrographs of SW480 and SW620 cells are shown with actin in green and cell nuclei in blue. In order to establish cytoskeleton organization differences among different cell lines, two parameters were considered: actin fibers Coherency and density of junctions of actin network (ρJ). Coherency is calculated from the structure tensor of each pixel in the image and is bounded between 0 (isotropic areas) and 1 (highly oriented structures) [10]. In Fig. 1.4a–c three representative confocal images of skeletonized actins filaments Fig. 1.3 Representative confocal images of Phalloidin labeled actin in SW480 and SW620 cells. Both elongated and rounded cells are visible in the microscope field in SW480 sample (left), while only rounded cells are visible in SW620 micrograph (right). Cells nuclei are labelled in blue withDAPI Fig. 1.4 Representative confocal images of skeletonized actin in SW480 E-type (a), SW480 R-type (b) and SW620 cells (c). Skeleton branches are labelled in orange, junctions in purple and end points in blue. On the right, skeletons are superimposed on the original confocal images of SW480 E-type (d), SW480 R-type (e) and SW620 (f) cells. Scale Bar is 22μmin (d, e) and10μmin (f). In theinsets detail of cytoskeleton of each cell are shown 4 V. Palmieri et al.
RkJQdWJsaXNoZXIy MTMzNzEzMQ==