Dapi fitc tritc

The fixed MRC-5 cells were examined using a wide-field inverse fluorescence microscope Olympus IX-81 (Olympus, Japan; xCellence software). The samples were monitored at 100 × magnification using a 10 × objective with a numerical aperture of 0.30. For fluorescence, a triple, quad filter DAPI/FITC/TRITC (Olympus, Japan) and a high-stability 150-W xenon arc burner (100 % intensity) were utilized. From each sample, at least ten regions of interest were taken, and the images were captured by an EM-CCD camera C9100-02 (Hamamatsu, Japan). Then, the fluorescence images were background-corrected, and the two fluorescence channels (DAPI and FITC) were merged.

The fixed samples of U-2 OS cells were examined by wide-field fluorescence microscopy. For this, an inverse Olympus IX-81 microscope (Olympus, Tokyo, Japan) with xCellence software was used. The stained cells were captured by 10×, 20×, and 40× objectives with the NAs of 0.30, 0.45, and 0.60, respectively. F-actin and cell nuclei were monitored using a triple quad filter DAPI/FITC/TRITC (Olympus, Tokyo, Japan) and the regions of interest were captured by an EM-CCD camera (Hamamatsu, Honshu, Japan). The fluorescence images were deconvolved (50%), background-corrected, and the fluorescence channels (DAPI and TRITC) were merged, when possible, i.e., for PS samples and controls. Since PES and PEN exhibit high background autofluorescence in the DAPI channel, it was not possible to efficiently capture cell nuclei of cells growing on these two materials. Then, the cell number was counted using Image J software 1.53.

The fixed and stained VAMPIRO U-2 OS cell samples were subjected to wide-field fluorescence microscopy using an inverse fluorescence microscope Olympus IX-81 (Olympus, Tokyo, Japan) operated with xCellence software. The fluorescently labeled cells were examined at 100× (10× objective, NA of 0.30), 200× (20× objective, NA of 0.45), and 400× (40× objective, NA of 0.60) magnification. Cell cytoplasm, cytoskeleton, and nuclei were imaged by a triple quad filter DAPI/FITC/TRITC (Olympus, Tokyo, Japan). The images were captured by an EM-CCD camera (Hamamatsu, Honshu, Japan). Then, the fluorescence images were background-corrected, deconvolved (50%), and all three fluorescence channels (DAPI, FITC, TRITC) were merged.