As new imaging methods are developed and our knowledge of neural development deepens, methods for growing neuronal cells in culture where they are readily manipulated and observed become more and more important. In the August issue of Cold Spring Harbor Protocols, Amy MacDermott and colleagues from Columbia University provide a series of protocols describing neuronal culture techniques that are particularly useful for studying synapse formation and interconnection.

Preparation of Coverslips for Neuronal Cultures describes setting up glass coverslips for three different types of cell culture: Mass Culture, which results in an extensive and widespread network of synapses, Microisland Culture, which allows easier identification of synaptically connected neurons, and Macroisland Culture, which strikes a balance between the better survival rates and better identification of synapses offered by the two methods above.

Survival of CNS neurons in culture is usually greatly improved if the neurons are plated on top of a confluent astrocyte layer, as is described in Dissection, Plating, and Maintenance of Cortical Astrocyte Cultures. Under these conditions, neurons attach more firmly and develop a more easily identifiable bi-dimensional neuritic tree than in culture conditions where primary cells are plated on top of collagen, laminin or other cell-free substrates.

Once established the astrocyte layer can be used to in the Dissection, Plating, and Maintenance of Dorsal Horn Neuron Cultures, which are suitable for electrophysiological, molecular and immunocytochemical studies. Dorsal horn neurons can be grown by themselves, or co-cultured with embryonic dorsal root ganglion neurons as described in Dissection, Plating, and Maintenance of Dorsal Root Ganglion Neurons for Monoculture and for Coculture with Dorsal Horn Neurons.