Fluorescence Proteins and Time-Lapse Imaging of the Cytoskeleton

In this chapter, we provide a detailed description for time-lapse imaging of microtubules, neurofilaments, and microfilaments within primary neurons. We use two primary neuronal cell culture systems for the analysis of different aspects of cytoskeletal motion and organization: (1) dissociated dorsal root ganglia, which are highly practical to study cytoskeletal dynamics along their neurites or within the growth cone, and (2) cerebellar slice cultures, which are characterized by their organotypic morphology even after 30 days in vitro. In particular in these slice cultures Purkinje cells exhibit highly dynamic dendritic spines within a functional neuronal network.