Human Lung Carcinoma Cells (A-549 Line)

In order to visualize the relationship between mitochondria and the cytokeratin intermediate filament network, a culture of human lung carcinoma (A-549) cells was immunofluorescently labeled with primary anti-cytokeratin (pan) mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Marina Blue. Mitochondria were simultaneously targeted with MitoTracker Red CMXRos and 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.