Monoclonal anti vinculin fitc

The cell morphology and cell spreading of MG63 osteoblasts on non-porous and porous PCL scaffolds with or without loaded DXM were evaluated by confocal laser scanning microscopy. MG63 osteoblasts were seeded on scaffolds at 50,000 cell/mL (approximately 500 cells/mm² of substrate) in complete culture media, and incubated for 1, 7, 14 and 21 days at 37 °C in 5% CO₂, 95% relative humidity. At the end of the incubation time, the non-attached cells were removed by rinsing carefully three times with PBS 1x. Focal adhesion contacts (FAC) were detected by cell staining with a monoclonal anti-vinculin-FITC (working dilution of 1:50, Sigma-Aldrich) according the manufacturer’s instructions. Cytoskeleton morphology was investigated by cell staining with phalloidin-TRITC at a final concentration of 1 mg/mL (Sigma-Aldrich), according the manufacturer’s instructions. Nuclei were stained with DAPI (Sigma-Aldrich). Confocal micrographs were taken with a Leica confocal scanning system mounted into a Leica TCS SP5 (Leica Microsystem GmbH, Mannheim, Germany), equipped with a 40x and 63x oil immersion objective and a spatial resolution of approximately 200 nm in x-y and 100 nm in z.