Preparation of chemotactically competent Dictyostelium cells

Abstract

Environmental changes result in signaling events in cell membranes. To develop the means to understand these events at the molecular level, it is essential to become familiar with the stochastic nature of signaling molecules in living cells. Using total internal reflection fluorescent microscopy (TIRFM), these signaling events can be directly observed at the single-molecule level. This protocol describes the preparation of chemotactically competent Dictyostelium cells. These cells are highly sensitive to chemoattractant stimulation and can be used to visualize single molecules with TIRFM.