Guinea Pig Colorectal Adenocarcinoma Epithelial Cells (GPC-16 Line)
The proximity of intermediate filaments and the cytoskeletal filamentous actin network was visualized by treating the fixed and permeabilized culture of guinea pig adenocarcinoma cells (GPC-16 line) presented above with mouse anti-vimentin primary antibodies followed by goat anti-mouse secondary antibodies (IgG) conjugated to Cy3. F-actin was subsequently labeled with Coumarin conjugated to phalloidin, and the nuclei were counterstained with SYTOX Green. Images were recorded in grayscale with a 12-bit digital camera coupled to either a Nikon E-600 or Eclipse 80i microscope equipped with bandpass emission fluorescence filter optical blocks. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.