Electron Microscopy Methods and Protocols

About this book

In this revised and expanded second edition, Electron Microscopy: Methods and Protocols presents the newest technology in electron microscopy, while maintaining the practicality and accessibility of the acclaimed first edition. Like the first edition, this volume provides clear, concise instructions on processing biological specimens and includes discussion on the underlying principles of the majority of the processes presented. Each protocol is presented by experts with years of first-hand experience and in-depth knowledge of the techniques they describe. The Notes section accompanying each protocol will prove invaluable for adapting and troubleshooting protocols. Electron Microscopy: Methods and Protocols, Second Edition, comprises two major areas of electron microscopy-transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The TEM area covers several key techniques, including: conventional specimen preparation methods for cultured cells and biomedical and plant tissues; cryospecimen preparation by high-pressure freezing and cryoultramicrotomy negative staining and immunogold labeling techniques; and TEM crystallography and cryo-TEM tomography. The SEM area similarly attends to conventional-, variable pressure-, environmental-, and cryoscanning microscopy techniques, as well as the application of X-ray microanalysis. Protocols for the application of X-ray microanalysis to SEM and mass spectrometry conclude the volume.

Contents

General Preparation of Material and Staining of Sections Heather Davies. Negative Staining of Thinly Spread Biological Particulates J. Robin Harris. Preparation of Thin-Film Frozen-Hydrated Vitrified Biological Specimens for Cryoelectron Microscopy J. Robin Harris and Marc Adrian. The Production of Cryosections Through Fixed and Cryoprotected Biological Material and Their Use in Imunocytochemistry Paul Webster. High Pressure Freezing for Preservation of High-Resolution Fine Structure and Antigenicity for Immunolabelling Kent McDonald. The Application of LR Gold Resin for Immunogold Labelling Julian Thorpe. Low- Temperature Embedding in Acrylic Resins P. Gounon. Quantitative Aspects of Immunogold Labelling in Embedded and Non-embedded Sections C.Rabouille. Microwave Processing Techniques for Electron Microscopy: A Four-Hour Protocol R. T. Giberson. Electron Microscopic Enzyme Cytochemistry Nobukazu Araki and Tanenori Hatae. In Situ Molecular Hybridization Techniques for Ultrathin Sections Jean-Guy Forunier. Application of Electron Microscopy for Morphological Study and Immunogold Localization of Proteins in both Wild Type and Mutant Yeast Strains Nasser Hajibagheri. Dark-Field Visualization of ds DNA and ss DNA Molecules by Electron Microscopy Nasser Hajibagheri. Applications of EM for Studying Protein-DNA Complexes Ross B. Inman. X-ray Miocroanalysis Techniques John Morgan.

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