Analytical tools to monitor exocytosis: a focus on new fluorescent probes and methods.

A great deal of research has been focused on unraveling the processes governing the exocytotic pathway and the extent of release during the process. Arguments abound for and against both the occurrence and significance of full release during exocytosis and partial release including kiss-and-run events. Several optical methods to directly observe the exocytosis process have been developed and here we focus on fluorescence methods and probes for this work. Although fluorescence imaging has been used for cell experiments for decades, in the last two decades a plethora of new approaches have arrived on the scene. These include application of new microscopy techniques, like total internal reflectance and stimulated emission depletion that are offering new ways to circumvent the limits of far field microscopy with a diffraction limit of 200 nm, and allow tracking of single synaptic vesicles. For selective imaging of synaptic vesicles the introduction of methods to stain the vesicular compartment has involved developing probes of the vesicular membrane and intravesicular solution, nanoparticle quantum dots that can be observed during exocytosis but not via the fusion pore, and fluorescent false neurotransmitters.