Celld analysis software

Actin labeling was performed in order to evaluate qualitatively F-actin microfilaments of cytoskeleton (involved in mobility and intracellular scaffold formation). Additionally, Hoechst staining was carried out to determinate the number of viable cells. Cells on the films were fixed at 96 h with 4% paraformaldehyde (PFA) solution for 10 min. After PFA was removed, cells were rinsed with PBS twice and permeabilized with 0.1% (v/v) Triton X-100. Then, the cells were washed with PBS and stained with Texas Red®-X phalloidin (Life Technologies), a highaffinity F-actin probe conjugated to red fluorochrome, for 20 minutes at room temperature and in darkness, followed by Hoechst staining (Invitrogen, Molecular Probes®). Finally fluorescent-labeled cells were observed using an inverted fluorescence microscope (Olympus IX51) with a TRICT filter (λ ex /λ em =550/600 nm) for Actin and DAPI filter for Hoechst (λ ex /λ em = 380/455 nm) using CellD analysis software (Olympus).

Cells were fixed with 4% paraformaldehyde (PFA) solution for 15 min. After PFA was removed, cells were rinsed with PBS twice and permeabilized with 0.1% (v/v) Triton X-100, washed with PBS and stained with Texas Red®-X phalloidin (Life Technologies), a high-affinity F-actin probe conjugated to red fluorochrome, for 20 min at room temperature and in darkness, followed by Hoechst staining (Invitrogen, Molecular Probes ®) for nuclei visualization. Finally fluorescent-labelled cells were observed using an inverted fluorescence microscope (Olympus IX51) with a TRICT filter (λex/λem = 550/600 nm) for Actin and DAPI filter for Hoechst (λex/λem = 380/455 nm) using CellD analysis software (Olympus).