Time-lapse imaging of membrane traffic in living cells

Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments--including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes--that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)-fusion proteins and recent advances in fluorescent microscope imaging systems. This protocol describes the use of the confocal laser-scanning microscope (CLSM) for time-lapse imaging of one or more fluorescent markers.