Embryonic Swiss Mouse Fibroblast Cells (3T3 Line)

The log phase culture of 3T3 cells illustrated above was fixed, permeabilized, blocked with 10-percent normal goat serum, and then treated with mouse anti-NPCP (nuclear pore complex proteins) primary antibodies followed by secondary antibodies conjugated to Alexa Fluor 568. The secondary cocktail contained Alexa Fluor 488 conjugated to phalloidin to label the filamentous actin network, and the nuclei were counterstained with Hoechst 33342. 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.