Electron Microscopy Methods and Protocols

1. Front Matter

   Pages i-xi

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2. General Preparation of Material and Staining of Sections

   • Heather A. Davies

   Pages 1-11

3. Negative Staining of Thinly Spread Biological Particulates

   • J. Robin Harris

   Pages 13-30

4. Preparation of Thin-Film Frozen-Hydrated/ Vitrified Biological Specimens for Cryoelectron Microscopy

   • J. Robin Harris, Marc Adrian

   Pages 31-48

5. The Production of Cryosections Through Fixed and Cryoprotected Biological Material and Their Use in Immunocytochemistry

   • Paul Webster

   Pages 49-76

6. High-Pressure Freezing for Preservation of High Resolution Fine Structure and Antigenicity for Immunolabeling

   • Kent McDonald

   Pages 77-97

7. The Application of LR Gold Resin for Immunogold Labeling

   • J. R. Thorpe

   Pages 99-110

8. Low-Temperature Embedding in Acrylic Resins

   • Pierre Gounon

   Pages 111-124

9. Quantitative Aspects of Immunogold Labeling in Embedded and Nonembedded Sections

   • Catherine Rabouille

   Pages 125-144

10. Microwave Processing Techniques for Electron Microscopy: A Four-Hour Protocol

    • Rick T. Giberson, Richard S. Demaree Jr.

    Pages 145-158

11. Electron Microscopic Enzyme Cytochemistry

    • Nobukazu Araki, Tanenori Hatae

    Pages 159-165

12. In Situ Molecular Hybridization Techniques for Ultrathin Sections

    • Jean-Guy Fournier, Françoise Escaig-Haye

    Pages 167-181

13. Preparation of the Fission Yeast Schizosaccharomyces pombe for Ultrastructural and Immunocytochemical Study

    • M.A. Nasser Hajibagheri, Kenneth Sawin, Steve Gschmeissner, Ken Blight, Carol Upton

    Pages 183-207

14. Preparation of Double/Single-Stranded DNA and RNA Molecules for Electron Microscopy

    • M.A. Nasser Hajibagheri

    Pages 209-227

15. Applications of Electron Microscopy for Studying Protein-DNA Complexes

    • Maria Schnos, Ross B. Inman

    Pages 229-243

16. X-Ray Microanalysis Techniques

    • A. John Morgan, Carole Winters, Stephen Stürzenbaum

    Pages 245-276

17. Back Matter

    Pages 277-283

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Electron Microscopy Methods and Protocols is designed for the established researcher as a manual for extending knowledge of the field. It is also for the newcomer who wishes to move into the field. A wide range of applications for the examination of cells, tissues, biological macromolecules, molecular structures, and their interactions are discussed. We have tried to gather together methods that we consider to be those most generally appli- ble to current research in both cell and molecular biology. Each chapter c- tains a set of related practical protocols with examples provided by experts who have first-hand knowledge of the techniques they describe. The individual chapters are grouped according to similarities in their specimen preparation and methodology. Methods are presented in detail, in a step-by-step fashion, using reproducible protocols the authors have personally checked. During the last decade, the scientific literature describing the use of colloidal gold as an immunocytochemical marker has increased at an ex- nential rate, and this trend is expected to continue. We have included a large number of variations on the immunogold labeling technique. In both the ne- tive staining and cryo chapters, authors emphasize the “immunological app- cations” in order to correlate as fully as possible with the emphasis on immunogold labeling in the other chapters. Electron Microscopy Methods and Protocols commences with the routine preparation of biological material for classical transmission electron microscopy involving tissue fixation, embedding, and sectioning (Chap. 1).

"Chapter after chapter, methods are described in detail following a step-by-step schedule giving sufficient details to lead the researcher to precise and reproducible work. . .One of the great features of this book is a series of notes concluding every chapter. These notes pool a lot of important recommendations. If I can give a piece of good advice, just for these notes, buy this book and read all this valuable information."-Biochemie

"This ring-bound volume provides the reader with a useful source of comprehensive and accurate protocols for electron microscopic specimen preparation with an emphasis on the subcellular level..."-Microbiology Today

• Imperial Cancer Research Fund, London, UK

  M. A. Nasser Hajibagheri

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