Fluorescein isothiocyanate fitc labeled phalloidin green fluorescence

The cytoskeleton was assessed by a confocal laser scanning microscope (Olympus FluoView FV1000). After culturing for 6 h, the cells on titanium samples were fixed in 3.7% formaldehyde for 5 min and then permeabilized with 0.1% Triton X-100. Subsequently, the samples were stained with 50 mg/mL fluorescein isothiocyanate (FITC)-labeled phalloidin (green fluorescence) (Sigma-Aldrich Co.) for 40 min, and then, nuclei were counterstained with DAPI (blue fluorescence) (Sigma-Aldrich Co.) for 5 min at room temperature in the dark. Finally, stained cells were inspected by confocal laser scanning microscopy (CLSM).

The cell morphology and details of cell/biomaterial interaction were inspected using FE-SEM and a confocal laser scanning microscope (CLSM) (Olympus FluoView FV1000). After incubation for 3 days, the samples with attached cells were fixed in 3% glutaraldehyde, dehydrated in a graded ethanol series, freeze dried, sputter coated with gold, and observed using FE-SEM.
After culturing for 3 days, the cells on the Ti samples were fixed in 3.7% formaldehyde for 5 minutes and then permeabilized with 0.1% Triton X-100. Subsequently, they were stained with 50 mg/mL fluorescein isothiocyanate (FITC)-labeled phalloidin (green fluorescence) (Sigma-Aldrich Co.) for 40 minutes and then counterstained with DAPI for the cell nuclei (blue fluorescence) (Sigma-Aldrich Co.) for 5 minutes at room temperature in the dark. Finally, the stained cells were inspected with a CLSM.