Recorded 18 November 2022. Alberto Bartesaghi of Duke University presents "High-throughput cryo-ET: from data acquisition to high-resolution structures" at IPAM's Cryo-Electron Microscopy and Beyond Workshop. Abstract: Tomographic reconstruction of frozen-hydrated specimens followed by extraction and averaging of sub-tomograms has successfully been used to determine the structure of macromolecules in their native environment at resolutions that are high enough to reveal molecular level interactions. The advent of high-throughput methods for data acquisition that can produce thousands of high-quality tilt series during a single microscope session, however, has uncovered bottlenecks in the downstream data analysis which has so far relied on supervised, user-driven pipelines. In this talk, I will present recent advances in high-throughput tomography that have allowed us to streamline the cryo-ET structure-determination process and improve the resolution of structures using the "constrained single-particle tomography" paradigm. The combination of strategies for accelerated tilt-series acquisition together with data-driven techniques for high-resolution image analysis, will pave the way for cryo-ET to become the technique of choice to determine protein structures in their native environment. Learn more online at: http://www.ipam.ucla.edu/programs/workshops/workshop-iii-cryo-electron-microscopy-and-beyond/?tab=schedule