Analyzing Structural Plasticity of Dendritic Spines in Organotypic Slice Culture

Understanding the rules of synapse dynamics in the context of development, learning, and nervous system disorders is an important part of several fields of neuroscience. Despite significant methodological advances, observations of structural dynamics of synapses still present a significant experimental challenge. In this chapter we describe a set of techniques that allow repetitive observations of synaptic structures in vitro in organotypic cultures of rodent hippocampus. We describe culturing of slices, transfection with reporter-carrying plasmids, repetitive imaging of dendritic spines with confocal laser scanning microscopy and analysis of spine morphology dynamics.