Cellular Electron Microscopy

Recent advances in the imaging technique electron microscopy (EM) have improved the method, making it more reliable and rewarding, particularly in its description of three-dimensional detail. Cellular Electron Microscopy will help biologists from many disciplines understand modern EM and the value it might bring to their own work. The book’s five sections deal with all major issues in EM of cells: specimen preparation, imaging in 3-D, imaging and understanding frozen-hydrated samples, labeling macromolecules, and analyzing EM data. Each chapter was written by scientists who are among the best in their field, and some chapters provide multiple points of view on the issues they discuss. Each section of the book is preceded by an introduction, which should help newcomers understand the subject. The book shows why many biologists believe that modern EM will forge the link between light microscopy of live cells and atomic resolution studies of isolated macromolecules, helping us toward the goal of an atomic resolution understanding of living systems.

Updates the numerous technological innovations that have improved the capabilities of electron microscopyProvides timely coverage of the subject given the significant rise in the number of biologists using light microscopy to answer their questions and the natural limitations of this kind of imagingChapters include a balance of "how to", "so what" and "where next", providing the reader with both practical information, which is necessary to use these methods, and a sense of where the field is going

Introduction: McIntosh - A new era in electron microscopy of cells

Preparative Methods for Cellular Electron Microscopy
1. The physics of rapid cooling and its implications for cryo-immobilization of cells
2. Cryopreparation Methods for Electron Microscopy of Selected Model Systems
3. Cryo-preparation methodology for plant cell biology
4. Correlative Light and Electron Microscopy of Early C. elegans Embryos in Mitosis
Imaging Fixed Cells in Three Dimensions
5. Understanding microtubule organizing centers by comparing mutant and wild-type structures with electron tomography
6. Whole Cell Investigation of Microtubule Cytoskeleton Architecture by Electron Tomography
7. Electron Microscopy of archaea
8. Reconstructing Mammalian Membrane Architecture by Large Area Cellular Tomography
9. Visualization of Membrane-Cytoskeletal Interactions during Plant Cytokinesis
10. Electron Tomographic Methods for Studying the Chemical Synapse
11. Using electron microscopy to understand functional mechanisms of chromosome alignment on the mitotic spindle
12. EM Analysis of the Leading Edge in Migrating Cells
13. Imaging actomyosin in situ

Imaging Frozen-Hydrated Cells and Cell Parts
14. Electron tomography of bacterial chemotaxis receptor assemblies
15. How to “read” a vitreous section
16. Single Particle Electron Cryomicroscopy of the Ion Channels in the Excitation-Contraction Coupling Junction
17. Electron microscopy of microtubule-based cytoskeletal machinery
18. Reconstructing the endocytotic machinery

Localizing Macromolecules in Cells
19. 3-D Immuno-localization with plastic sections
20. EM analysis of viral morphogenesis
21. Electron tomography of immuno-labeled cryosections
22. Visualizing macromolecules with fluoronanogold: from photon microscopy to electron tomography
23. Markers for correlated light and electron microscopy
24. Localizing Specific Elements Bound to Macromolecules by EFTEM
25. Localization of protein complexes by pattern recognition

Aspects of Data Collection and Analysis Instrumentation for image acquisition
26. Application of Energy Filtered Electron Microscopy to Tomography of Thick, Selectively Stained Biological Samples
27. Optimization of Image Collection for Cellular Electron Microscopy
28. Future Directions for Camera Systems in Electron Microscopy

Computational methods for the analysis of cellular data
29. Structure determination in situ by averaging of tomograms
30. Methods for image segmentation in cellular tomography
31. Database Resources for Cellular Electron Microscopy