Ix50 inverted system microscope

Cells directly attached onto Matrigel-coated coverslips (BD Biosciences), were fixed with 1% paraformaldehyde (Sigma-Aldrich) in PBS containing 0.1% Triton X-100. Cells were washed three times with PBS and incubated with the corresponding antibodies and 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich). After incubation, samples were mounted with Mounting Fluorescence Medium (Dako), and images were captured using a TCS-SP2-AOBS-UV confocal microscope (Leica) with a ×63 oil-immersion objective. Displayed images were captured at the same sections in the different samples. Bright-field images were obtained using an IX50 inverted system microscope (Olympus), with LCPLFL ×20 and ×40 objectives. Cell and nucleus diameter and area were measured using ImageJ software.