In a double immunofluorescence experiment, a coronal rat hippocampus tissue section was labeled for the neuron-specific class III beta-tubulin isoform and heavy chain neurofilament subunits by treating the specimen with a cocktail of mouse anti-beta-III-tubulin and chicken anti-NF-H primary antibodies followed by goat anti-mouse and anti-chicken secondary antibodies (IgG) conjugated to Alexa Fluor 488 and Alexa Fluor 568, respectively. Nuclear DNA was counterstained with DRAQ5. Images were recorded with a 40x oil immersion objective using a zoom factor of 1.2 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, the Alexa Fluor 488 image channel was pseudocolored red, the Alexa Fluor 568 channel was pseudocolored yellow, and the DRAQ5 channel was pseudocolored cyan. View a larger version of this digital image. |