Cryo-Electron Tomography and Automatic Segmentation of Cultured Hippocampal Neurons

Cryo-electron tomography is fast becoming a preferred method for studying intracellular environments at the molecular scale. Increases in data collection throughput means that large numbers of tomograms can be generated at rates too fast for humans to easily explore quantitatively. Currently, there is a large effort to make data collection and segmentation tools more automated. Here, we describe a workflow for preparing cultured neurons on electron microscopy grids, batch tomographic data collection, reconstruction and automatic segmentation using freely and commercially available software.