The mode of cell migration was originally classified based on the morphology of migration patterns. This terminology was then extended to include molecular parameters, such as cytoskeletal organization. As main categories, cell move either individually (amoeboid or mesenchymal) or collectively (the migration of cohesive multicellular units. The single cell migration is an important part of researches on cell migration and many methods can be used to research it. Among all the methods, Scanning electrochemical microscopy (SECM) is a unique and powerful technique to study single cell migration, which can detect the single-cell movement in three-dimensional (3D) tissue environments.