Male Rat Kangaroo Kidney Epithelial Cells (PtK2 Line)
After treatment for one hour with MitoTracker Red CMXRos in serum-based growth medium, the adherent culture of PtK2 cells presented above was fixed in 50:50 methanol:acetone, permeabilized, blocked, and treated with mouse anti-cytokeratin (pan) monoclonal antibodies. The intermediate filaments were visualized with goat anti-mouse secondary antibodies (IgG) conjugated to fluorescein, and the nuclei were counterstained with DAPI. 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.