Dopamine is a neurotransmitter critical in mediating reward pathways, locomotion, and learning and memory. Dopamine visualization is crucial for studying dopamine localization, release, and subcellular quantitation in the brain. We have developed a new immunostaining protocol to directly detect dopamine (DA) in brains of Drosophila melanogaster with high signal to noise ratio. The DA antibody stains dopamingergic neurons and terminal projections in wild-type flies while DA deficient brains lack staining, showing specificity of the antibody. The common method to infer DA in the brain utilizes immunostaining of tyrosine hydrolyase (TH), the rate limiting enzyme for DA synthesis. Here, we show the flaw in using this marker, since detectable levels of TH produced by poorly translated TH protein can produce normal amounts of dopamine. Reduced DA staining in the terminal region of a vesicular transporter mutant tell us that stained dopamine is contained in vesicles. This new immunodetection protocol will allow accurate and direct visualization of changes in subcellular localization of DA as a function of other genetic and behavioral state alterations.