2024 Cryo-Electron Tomography Data Processing Workshop

Cryo-electron tomography is a rapidly evolving structural technique used to calculate 3D structures of biological specimens in vitro and within cells, i.e., in situ. The power of this approach is its ability to chart native cellular landscapes at high resolutions. Technological developments in microscopes, cameras, and computation make it possible to determine sub-10 Å 3D structures at molecules directly within cells. 

This four-day workshop will introduce participants to the image processing work-flow used to analyze tilt-series data and mine its structural information. Working with instructors, participants will learn how to independently use software packages such as Warp/M, IMOD, STOPGAP, tomoDRGN and Amira. The course will cover all aspects of cryo-ET data processing, including tilt-series alignment, tomogram reconstruction, particle picking, subtomogram averaging, and segmentation. The class will use the in situ ribosome dataset (Tegunov et al., Nature Methods, 2021) for all processing steps except for segmentation. Segmentation protocols will be taught using a mammalian cell tomogram provided to all participants.

Participant Requirements:

• Laptop computer
• Watch Dr. Jensen's Caltech online cryo-EM course before arriving
• Experience collecting cryo-ET data and familiarity with the SerialEM data format will be beneficial.