Deep Learning-Based Segmentation of Cryo-Electron Tomograms

Jessica E. Heebner, Carson Purnell, Ryan K. Hylton, Mike Marsh, Michael A. Grillo, Matthew T. Swulius

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


Cryo-electron tomography (cryo-ET) allows researchers to image cells in their native, hydrated state at the highest resolution currently possible. The technique has several limitations, however, that make analyzing the data it generates time-intensive and difficult. Hand segmenting a single tomogram can take from hours to days, but a microscope can easily generate 50 or more tomograms a day. Current deep learning segmentation programs for cryo-ET do exist, but are limited to segmenting one structure at a time. Here, multi-slice U-Net convolutional neural networks are trained and applied to automatically segment multiple structures simultaneously within cryo-tomograms. With proper preprocessing, these networks can be robustly inferred to many tomograms without the need for training individual networks for each tomogram. This workflow dramatically improves the speed with which cryo-electron tomograms can be analyzed by cutting segmentation time down to under 30 min in most cases. Further, segmentations can be used to improve the accuracy of filament tracing within a cellular context and to rapidly extract coordinates for subtomogram averaging.

Original languageEnglish (US)
Article numbere64435
JournalJournal of Visualized Experiments
Issue number189
StatePublished - Nov 2022

All Science Journal Classification (ASJC) codes

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology


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