Time-Lapse Imaging of Membrane Traffic in Living Cells

Abstract

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.