A semi-confluent culture of A-10 cells (illustrated above) was fixed, permeabilized, and blocked with 10-percent normal goat serum in phosphate-buffered saline prior to immunofluorescent labeling with primary antibodies to giantin, a protein resident in the Golgi complex of mammalian cells. The culture was subsequently stained with a mixture of secondary antibodies conjugated to Alexa Fluor 568 in a mixture containing phalloidin conjugated to Alexa Fluor 488 (pseudocolored blue). The cell nuclei were counterstained with the red-absorbing dye TO-PRO-3 (pseudocolored green). Images were recorded with a 60x oil immersion objective using a zoom factor of 3.0 and sequential scanning with the 488-nanometer spectral line of an argon-ion laser, the 543-nanometer line from a green helium-neon laser, and the 633-nanometer line of a red helium-neon laser. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles unless otherwise noted above. View a smaller version of this digital image. |