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.
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