Smart parallel automated cryo-electron tomography.

Journal: Nature methods
PMID:

Abstract

In situ cryo-electron tomography enables investigation of macromolecules in their native cellular environment. Samples have become more readily available owing to recent software and hardware advancements. Data collection, however, still requires an experienced operator and appreciable microscope time to carefully select targets for high-throughput tilt series acquisition. Here, we developed smart parallel automated cryo-electron tomography (SPACEtomo), a workflow using machine learning approaches to fully automate the entire cryo-electron tomography process, including lamella detection, biological feature segmentation, target selection and parallel tilt series acquisition, all without the need for human intervention. This degree of automation will be essential for obtaining statistically relevant datasets and high-resolution structures of macromolecules in their native context.

Authors

  • Fabian Eisenstein
    Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
  • Yoshiyuki Fukuda
    Institute of Advanced Medical Sciences, Tokushima University, Tokushima, Japan.
  • Radostin Danev
    Graduate School of Medicine, University of Tokyo, Tokyo, Japan. rado@m.u-tokyo.ac.jp.