Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s disease. Among multiple pathological features, the loss of dopaminergic neurons of substantia nigra is the most representative one. However, the molecular details of the loss have not been clearly understood. In this work, we demonstrate the super-resolution, multiplexed, volumetric, three-dimensional imaging of dopaminergic neurons and other associated cells of substantia nigra of the brain via expansion microscopy. To achieve multiplexed, volumetric, and super-resolution imaging of the brain, we screen multiple fluorophores and use the ones yielding the highest brightness. Super-resolution volumetric imaging of dopaminergic neurons and associated cells of the brain would help us to better understand the molecular mechanisms of PD.