HaloTag as a tool to investigate peroxisome dynamics in cultured mammalian cells

Peroxisomes are multifunctional organelles that can rapidly modulate their morphology, number, and function in response to changing environmental stimuli. Defects in any of these processes can lead to organelle dysfunction and have been associated with various inherited and age-related disorders. Progress in this field continues to be driven by advances in live-cell imaging techniques. This chapter provides detailed protocols for the use of HaloTag to fluorescently pulse-label peroxisomes in cultured mammalian cells. In contrast to the use of classical fluorescent proteins, this technology allows researchers to optically distinguish pools of peroxisomal proteins that are synthesized at different time points. The protocols can be easily adapted to image the dynamics of other macromolecular protein assemblies in mammalian cells.