Characterizing the detailed structure of the mammalian synapse is of crucial importance to understand its mechanisms of function. Here I describe a protocol to study synaptic architecture by cryo-electron tomography (cryo-ET), a powerful electron microscopy technique that enables 3D visualization of unstained, fully hydrated cellular structures at molecular resolution. The protocol focuses on purified synaptic terminals ("synaptosomes"), currently the most suitable preparation to analyze mammalian synaptic architecture by cryo-ET.
Keywords: Cryo-EM; Cryo-electron microscopy; Neuron; Postsynaptic density; Synaptic vesicle; Tissue fractionation.