Beschreibung:
This book presents key aspects and recent developments of cryogenic sample electron tomography (cryo-ET) methodology, authored by leading experts in the field. Understanding structure and function of biomolecules in the context of cells is a new frontier in cellular and structural biology. To facilitate such research, cryo-ET is a key method to visualize the molecules of life in their native settings. Cryo-ET enables the imaging of samples that are preserved in a near-native state, at (macro)-molecular resolution and in three dimensions. Thus, this technique is a unique tool to gain insights into how biomolecules collaborate in orchestrating fundamental biological processes, how mutations cause diseases, pathogens cause infections, and to develop novel therapeutics to treat such illnesses. This book provides a unique reference for the emerging field of cryo-ET.
1. Fundamentals of Instrumentation and electron optics For cryo-electron tomography.- 2. Principles of tomographic reconstruction .- 3. Cryo-preparation for cellular tomography.- 4. Cryo-FIB milling.- 5. In Situ Cryo-Electron Tomography and Advanced Micromanipulator Techniques.- 6. Labeling approaches and correlation with light microscopy (Cryo) CLEM.- 7. Data collection.- 8. Subvolume averaging and classification.- 9. High resolution tomography/ Molecular model building and visualization.- 10. Beam-induced Motion Mechanism and Correction for Improved Cryo-Electron Microscopy and Cryo-Electron Tomography.- 11. Tomogram segmentation.- 12. Electron tomography of cryo-fixed and resin-embedded samples.- 13. Democratization of tomographic data / FAIR data.