Using in situ cryoET to investigate intracellular host-pathogen interactions

In situ cryo-electron tomography can reveal interactions between pathogens and their hosts inside intact cellular environments. This seminar uses apicomplexan parasites and gram-negative bacteria to explain the workflow from sample preparation through image acquisition and analysis. It covers freezing, cryo-focused ion beam milling, cryo-electron tomography, correlative light and electron microscopy, tomogram reconstruction, segmentation and subtomogram averaging. The presentation discusses which aspects of the host-pathogen interface these approaches can resolve, their advantages and limitations, and when they suit a research question. Familiarity with the organisms or cryo-electron microscopy is helpful but not required.