Intracellular dopamine dynamics is a prelude to the sensation of reward and to motor control, and also a key to understanding substance addiction and Parkinson's disease. Dopamine neurotransmission has been investigated extensively, yet direct optical probing of dopamine has not been possible till now. Here we image intracellular dopamine with sub-micron three-dimensional resolution, by harnessing its auto-fluorescence with two-photon ultraviolet excitation (using a femtosecond optical parametric oscillator with output at 540 nm) and non-epifluorescent detection. The technique is established by first imaging dopamine in the dopaminergic cell line MN9D. These cells appear to be bright, and the signal strength reduces upon amphetamine administration (amphetamines are known to be powerful dopamine releasing agents). We then show that individual dopamine vesicles in the substantia nigra region can be imaged inside cultured brain slices. The cell brightness is much less in regions away from the nigra. Our technique can follow the intracellular events preceding dopamine release induced by depolarization and amphetamine exposure in these slices. This provides a unique assay for following any neurophysiological process that affects the intracellular dopamine dynamics.