Expression of genetically encoded fluorescent probes to monitor phospholipid dynamics in live neutrophils

Essential functions of neutrophils, including chemotaxis and phagocytosis, are directed in part by phospholipid signaling. Detailed elucidation of these pathways was hampered by the paucity of methods to study phospholipid localization and dynamics. The development of genetically encoded lipid-specific probes circumvented this limitation. The probes are chimeric constructs consisting of a specific lipid-binding domain fused to a fluorescent protein. This chapter describes a protocol to transiently transfect primary murine neutrophils with such probes in order to localize phospholipids in live cells, and provides a compendium of the types of lipid-binding domains currently used to visualize phospholipids.