Electron Microscopy in Microbiology

About this book

Electron Microscopy in Microbiology provides detailed descriptions of electron microscopic techniques relevant to all fields of microbiological research. In numerous protocols, standard techniques required for the structural characterization of unicellular organisms (suspended or fixed in microbial biofilms) and subcellular components (enzymes, nucleic acids, inclusion bodies, cell appendages etc) are described. Particular attention is given to techniques that provide information on the structure and compartmentation of biological macromolecules within the cell. These techniques include epitope mapping and localization of proteins by antibodies and other marker systems (DNA/RNA probes, lectins etc)

Contents

- Plastic support films
- Carbon and carbon-coated support films
- Preparation of hydrophilic films by glow discharge
- Other support films
- Negative staining
- Visualization of nucleic acids
- Sample preparation by rapid freezing
- Pretreatment of subcellular components
- Pretreatment of cells
- Freezing methods
- Air-dried or freeze dried, metal-shadowed specimens
- Freeze fracturing and freeze etching
- Sample preparation for thin sectioning
- Chemical fixation
- Dehydration, infiltration and embedding
- Ultramicrotomy
- Post-staining of sections
- Cryo-ultamicrotomy
- Localization of macromolecules
- Preparation of marker systems for localization procedures
- Localization procedures
- Control experiments in immunocytochemistry
- Labelling of epitopes on isolated protein molecules
- Imaging and image evaluation at the transmission electron microscope
- Stereo images
- Standard specimen preparation
- Preparation of a specimen for SEM
- Quantitation of results (surface area, particle counting)
- Densitograms
- Markham rotation

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