Cryo-Electron Tomography and Automatic Segmentation of Cultured Hippocampal Neurons

Ryan K Hylton; Victoria H Seader; Matthew T Swulius

doi:10.1007/978-1-0716-0966-8_2

Cryo-Electron Tomography and Automatic Segmentation of Cultured Hippocampal Neurons

Methods Mol Biol. 2021:2215:25-48. doi: 10.1007/978-1-0716-0966-8_2.

Authors

Ryan K Hylton¹, Victoria H Seader¹, Matthew T Swulius²

Affiliations

¹ Department of Biochemistry and Molecular Biology, The Penn State College of Medicine, Hershey, PA, USA.
² Department of Biochemistry and Molecular Biology, The Penn State College of Medicine, Hershey, PA, USA. mts286@psu.edu.

PMID: 33367998
DOI: 10.1007/978-1-0716-0966-8_2

Abstract

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.

Keywords: CET; Cryo; Cryo-EM; Cryo-ET; ECT; EMAN2; IMOD; Neural Network; Neuron; Segmentation; Tomography.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cells, Cultured
Cryoelectron Microscopy / methods*
Electron Microscope Tomography / methods*
Hippocampus / cytology*
Hippocampus / ultrastructure
Image Processing, Computer-Assisted
Imaging, Three-Dimensional
Neurons / cytology
Neurons / ultrastructure*
Rats
Software
Specimen Handling
Workflow