Cryo-electron Tomography for Cell Biology
On-demand
Dr. Ben Engel
Dr. Engel received his BS from UC-Berkeley and PhD from UC-San Francisco. His group at the Max Planck Institute of Biochemistry, applies in situ cryo-electron tomography (cryo-ET) to a wide variety of cellular questions to understand the relationship between the form of the organelle and the function of its resident macromolecules. By imaging the macromolecules within their native cellular environment and through a combination of nanometer-precision localization and high-resolution structural analysis, they aim to chart the molecular landscapes of organelles. His studies have been featured in journals including Cell, Nature, Science, PNAS, and eLife.
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In this tutorial, you will find:
- The complete workflow for in situ cryo-electron tomography
- How subtomogram averaging within the cell yields native-state structures of macromolecular complexes (e.g., the asymmetric and dilated nuclear pore of algae)
- How mapping these structures back into the native cellular environment reveals new molecular interactions that are only accessible by this technique (e.g., the binding of cargo to COPI-coated Golgi membranes and the tethering of proteasomes to the nuclear pore).
Cryo-electron tomography can visualize macromolecular structures in situ, inside the cell. Vitreous frozen cells are first thinned with a focused ion beam and then imaged in three dimensions using a transmission electron microscope. This transformative method has the power to revolutionize our understanding of cell biology, revealing native cellular architecture with molecular clarity.
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