Analysis of Random Migration of Dictyostelium Amoeba in Confined and Unconfined Environments.

Dictyostelium discoideum has proven to be an excellent model to study amoeboid cell migration. During their life cycle, Dictyostelium cells exhibit distinct modes of motility. Individual growth-phase cells explore new territories by random cell migration using the core cell motility machinery, but they can also hunt bacteria by detection and chemotaxis toward the by-product folate. After depletion of nutrients, the cells initiate a developmental program allowing streaming of the cells into aggregation centers by chemotaxis toward cAMP and by cell-to-cell adhesion. Subsequent development is associated with complex rotational movement of the compacted aggregates to drive cell type specific sorting, which in turn is necessary for terminal culmination and formation of fruiting bodies. Here we describe a protocol for the analyses of cell motility of vegetative Dictyostelium cells in unconfined and mechanically confined settings.