Propargylcholine bromide is a choline analogue containing terminal propargyl that can be incorporated into all classes of Choline-containing phospholipids such as phosphatidylcholine and sphingomyelin, labeling Choline-containing phospholipids. Propargylcholine bromide-labeled phospholipid molecules can be visualized in cells with high sensitivity and spatial resolution. Propargylcholine bromide can be used as a molecular tool to study the biochemical and metabolic processes of Choline-containing phospholipids in cells.