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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.
The topics covered range from the fundamental principles of imaging to sample preparation, data analysis,and data sharing within the scientific community. It serves as a valuable resource for the next generation of structural biologists, making it suitable both for undergraduate students studying biochemistry, biophysics, and molecular biology and highly valuable for the more experienced and specialized PhD student. Furthermore, it stands as a state-of-the-art source of knowledge for the established senior scientist within the field of structural biology.
List of contents
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.
About the author
Friedrich Förster has received his PhD from the Technical University Munich for his thesis work performed at the Max-Planck Institute of Biochemistry in Martinsried, Germany. He has spent 3 years as a postdcotoral scholar at the University of California in San Francisco, US; in 2009 he became independent research group leader at the Max-Planck Institute of Biochemistry, Germany and since 2016 Friedrich is Full Professor at the Department of Chemistry, University of Utrecht, The Netherlands. Friedrich Förster serves as the director of the Bijvoet Centre for Biomolecular Research.
Ariane Briegel received her PhD from the Technical University Munich for her thesis work performed at the Max-Planck Institute of Biochemistry in Martinsried,Germany. She spent 5 years as postdoctoral researcher and subsequent as a research scientist at the California Institute of Technology and the Howard Hughes Medical Center in the US. In 2015 Ariane became Full Professor in Ultrastructural Biology at the Leiden University, The Netherlands. Ariane Briegel is Co-Director of the Netherlands Center for Electron Nanoscopy,
