ABSTRACT
This chapter provides an overview of live-cell imaging techniques applied in laboratory to investigate the biosynthesis, the structure, and the function of gap junctions. It discusses microscope requirements, fluorescent probes, and imaging systems. The chapter also discusses how to image specific stages of the gap junction life cycle such as trafficking, endocytosis, and degradation. It describes how to make observations of the gap junction plaque structure and dynamics, the colocalization of gap junctions with their binding partners, and how to conduct quantitative fluorescence analysis of these observations. The chapter explains the techniques to allow for the imaging of gap junctions in living cells for several days without significant detrimental side effects. It point outs pitfalls and typical mistakes that can occur when conducting live-cell observations on gap junctions and give hints and tips on how to avoid them.
