View Tomo: Context-aware targeting and analysis in electron cryo-tomography
Gebauer, R.; Machala, E. A.; Mironova, Y.; Jönsson, M.-R.; Mazur, J.; Feldmann, C. A.; Zimmeck, M. A.; Silvester, E.; Caragliano, E.; Falckenhayn, J.; Yuen, E. L. H.; Ibrahim, T.; Hellert, J.; Bozkurt, T. O.; Kaufmann, R.; Quemin, E. R. J.; Grünewald, K.; Prazak, V.
Show abstract
Electron cryo-tomography (cryoET) resolves cellular structure in three dimensions, yet region selection is still typically based on two-dimensional projection images. Here, we introduce View Tomo, a workflow for rapid acquisition of low-magnification tomograms that enables screening, targeting and analysis in 3D. View Tomo tilt series are acquired in minutes at low dose (~3 e-/[A]2), producing high-contrast tomograms that remain compatible with subsequent high-resolution structural determination. We implemented View Tomo using an automated acquisition and reconstruction pipeline for rapid alignment. Across multiple viral and cellular systems, view tomograms revealed membrane remodelling events, assembly intermediates and cellular organisation that are difficult to identify in projection images. These data enabled targeted high-resolution imaging and quantitative analysis of spatial relationships within cells. View Tomo therefore extends cryoET workflows by improving target selection, enabling analysis of mesoscale organisation, and facilitating integration with correlative imaging approaches.
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