Madin-Darby Canine Kidney Epithelial Cells (MDCK Line)

Epithelial cell tight junctions and nuclear pore complex proteins were simultaneously imaged in a culture of Madin-Darby canine kidney cells with a cocktail of rabbit anti-ZO-3 and mouse anti-NPCP primary antibodies, followed by goat anti-rabbit and anti-mouse secondary antibodies conjugated to Alexa Fluor 568 and Alexa Fluor 488, respectively. 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.