Quantum dot-single-particle tracking (QD-SPT) is a super-resolution imaging technique that uses semiconductor nanocrystal quantum dots as fluorescent probes and is a powerful tool for analyzing protein and lipid behavior in the plasma membrane. Recent QD-SPT experiments have provided critical insight into the mechanism and physiological relevance of membrane self-organization in neurons and astrocytes in the brain. The mobility of some membrane molecules may become abnormal in cellular models of epilepsy and Alzheimer’s disease. Based on these findings, we propose that the behavior of membrane molecules reflects the condition of neurons in pathological disease states. In this chapter, we describe the latest, simple QD-SPT technique, which is feasible with epifluorescence microscopy and dissociated cell cultures.