Neurons in vitro are different from any other cell types in their sensitivity and complexity. Growing, differentiating, transfecting, and recording from single neurons and neuronal networks all present particular challenges. Some of the difficulties arise from the small scale of cellular structures, and have already seen substantial advances due to nanotechnology, particularly highly fluorescent semiconductor nanoparticles. Other issues have less obvious solutions, but the complex and often surprising way that novel nanomaterials react with cells have suggested some revolutionary approaches. We review some of the ways nanomaterials and nanostructures can contribute to in vitro neuroscience, with a particular focus on emphasizing techniques that are widely accessible to many laboratories and on providing references to protocols and methods. The issues of nanotoxicology of greatest interest to cultured neurons are discussed. Finally, we present some future trends and challenges in nano-neuroscience.