Use of fluorescence photobleaching techniques to measure the kinetics of intracellular transport

Transport into/out of subcellular compartments such as the nucleus is central to many important biological processes. In recent decades, significant advances in the development of fluorescent proteins has led to the development of various techniques to examine the movement/diffusion of proteins in living cells, enabling quantitative kinetic data to be generated and facilitating insight into the underlying molecular mechanisms. Here, we discuss the use of fluorescence photobleaching techniques such as fluorescence recovery after photobleaching (FRAP) to examine intracellular protein transport, including movement into and out of the nucleus, cytoplasmic/nuclear retention of proteins and lateral movement within membranes.