Human Fetal Lung Fibroblast Cells (MRC-5 Line)

In order to examine structural features of the Golgi complex and nucleus, a log-phase culture of MRC-5 cells (illustrated above) was fixed, permeabilized, blocked with normal goat serum, and then treated with rabbit anti-giantin (Golgi protein) primary antibodies followed by goat anti-rabbit secondary antibodies (IgG) conjugated to Alexa Fluor 568. Cell nuclei were counterstained with Hoechst 33258. 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.