Live Cell Imaging of the Twitching Motility of Cyanobacteria by High-Resolution Microscopy

Many cyanobacteria show directional movement either toward or away from light sources. The cell movement, also known as twitching motility, is usually driven by type IV pili (T4P), a bacterial molecular machine. The machine generates a propulsion force through repeated cycles of extension and retraction of pilus filaments. Here, I describe a phototaxis assay for observing Synechocystis sp. PCC6803 and Thermosynechococcus vulcanus at the single-cell level with optical microscopy. By adding fluorescent beads, I also describe a method how to visualize the asymmetric activation of T4P during phototaxis.