Front Cover -- BIOMEDICAL ELECTRON MICROSCOPY -- Copright Page -- FOREWORD -- PREFACE -- ACKNOWLEDGMENTS -- CHAPTER 1. MICROGRAPH INTERPRETATION -- 1. Classical Preparation Method -- 2. Low Temperature Approach -- 3. A Common Test Specimen -- 4. Detection of Objects -- 5. Identification of Artifacts -- 6. Analysis of Geometry -- 7. Biological Identification -- 8. Biological Diversity -- 9. Analysis of Dynamics: Endocytosis -- 10. Analysis of Dynamics: Synthesis -- 11. Comparison of Methods -- 12. Variations in Magnifications -- 13. Interpretation Difficulties -- 14. Diagnostic Pathology -- CHAPTER 2. FIXATIVES -- 1. Osmium Tetroxide and Glutaraldehyde at Low Magnification -- 2. Osmium Tetroxide and Glutaraldehyde at High Magnification -- 3. Glutaraldehyde Concentration: Perfusion Fixation -- 4. Glutaraldehyde Concentration: Immersion Fixation -- 5. Long Fixation Times -- 6. Formaldehyde-Glutaraldehyde Combinations -- 7. Potassium Permanganate, Picric Acid, and Ruthenium Red -- 8. Lead Salts and Tannic Acid -- 9. Uranyl Acetate Postfixation -- 10. Tannic Acid-Uranyl Acetate Variations -- 11. Osmium Tetroxide-Potassium Ferrocyanide -- 12. Osmium Tetroxide Artifacts -- 13. Glutaraldehyde Artifacts -- CHAPTER 3. FIXATIVE VEHICLE -- 1. Absence and Presence of Buffer -- 2. Comparison of Buffers -- 3. Osmolality of Perfusion Fixatives -- 4. Effects of Osmolality on Cell Shape -- 5. Effects of Osmolality on Cell Organelles -- 6. Adjustment of Osmolality with Sucrose -- 7. Colloid Osmotic Pressure: Low Magnification -- 8. Colloid Osmotic Pressure: High Magnification -- 9. Phosphate Buffer Precipitate -- CHAPTER 4. FIXATIVE APPLICATION -- 1. Perfusion-Fixation versus Immersion-Fixation -- 2. Perfusion-Fixation with Pressure Control -- 3. Fixation by Dripping in Vivo -- 4. Immersion-Fixation -- 5. Variability within the Tissue.

6. Unsuccessful Perfusion-Fixation -- 7. Superficial Tissue Damage -- 8. Early Postmortal Changes -- 9. Late Postmortal Changes -- 10. Influence of Biopsy Method -- 11. Microwave Treatment -- CHAPTER 5. DEHYDRATION AND EMBEDDING -- 1. Stepwise versus Direct Dehydration -- 2. Prolonged Dehydration in Ethanol -- 3. Prolonged Dehydration in Acetone -- 4. Inert Dehydration -- 5. Choice of Intermediate Solvent -- 6. Epon, Araldite, and Vestopal: Unstained Sections -- 7. Epon, Araldite, and Vestopal: Stained Sections -- 8. Different Brands of Epoxy Resins -- 9. Spurr and LR White -- 10. Embedding of Isolated Cells -- CHAPTER 6. FREEZING AND LOW-TEMPERATURE EMBEDDING -- 1. Plunge Freezing -- 2. Contact Freezing of Unfixed Tissue -- 3. Contact Freezing of Fixed Tissue -- 4. High-Pressure Freezing -- 5. Freeze-Substitution in Methanol/Uranyl Acetate -- 6. Freeze-Substitution in Osmium Tetroxide Acetone -- 7. Progressive Lowering of Temperature Embedding in Lowicryl -- CHAPTER 7. SUPPORT FILMS -- 1. Surface Topography -- 2. Stability of Film or Section -- 3. Holey Films -- 4. Thick and Thin Support Films -- 5. Folds in Support Film -- 6. Defects in Formvar Films -- 7. Common Contaminants -- 8. Volatile Contamination -- CHAPTER 8. ULTRAMICROTOMY -- 1. Correlation of Light and Electron Microscopy -- 2. Section Thickness: Low Magnification -- 3. Section Thickness: High Magnification -- 4. Section Thickness: Half-Micron Section -- 5. Determination of Section Thickness -- 6. Folds in the Section -- 7. Collection of Sections -- 8. Surface Topography of Sections -- 9. Knife Scratches -- 10. Mottling and Flaking -- 11. Worn Glass Knives -- 12. Transmitted Vibrations -- 13. Vibrations and Knife Marks -- 14. Selective Chatter -- 15. Compression -- 16. Holes and Deformations -- 17. Contamination during Microtomy -- 18. Extraction during Sectioning.

19. Cryoultramicrotomy: Survey Sections -- 20. Collection of Cryosections -- 21. Thickness of Cryosections -- 22. Staining of Cryosections -- 23. Defects in Cryosections -- CHAPTER 9. SECTION-STAINING -- 1. Lead Citrate Staining -- 2. Uranyl Acetate Staining -- 3. Enhanced Section Staining -- 4. Effects of Grid Storage -- 5. Section Exposed to Electron Beam -- 6. Effect of Electron Beam -- 7. Lead-Staining Granularity -- 8. Contamination -- 9. Block-Staining Precipitate -- 10. Removal of Contamination -- CHAPTER 10. MICROSCOPY -- 1. Resolving Power -- 2. Through-Focus Series: Hole and Latex Particle -- 3. Through-Focus Series: Myelin Sheath -- 4. Through-Focus Series: Cells -- 5. Minimum Contrast Focusing -- 6. Wobbler Focusing -- 7. Accelerating Voltages 20-100 kV -- 8. Accelerating Voltages 80-200 kV -- 9. Unsaturated Electron Beam -- 10. Condenser Apertures -- 11. Objective Aperture -- 12. Through-Focus Series: Astigmatism -- 13. Image Distortion -- 14. Chromatic Aberration -- 15. Mechanical Instability -- 16. Specimen Drift versus Astigmatism -- 17. Focus Drift -- 18. Electrical Instabilities -- 19. Contamination in the Electron Beam -- 20. Radiation Damage -- 21. Radiation Damage and Contamination -- 22. Low-Dose Exposure -- 23. Spectroscopic Imaging: Thin Film -- 24. Spectroscopic Imaging: Thick Section -- 25. Spectroscopic Imaging: Carbon -- 26. Spectroscopic Imaging: Calcium -- 27. Spectroscopic Imaging: Contrast Changes -- 28. Cryoelectron Microscopy: Na,K-ATPase Crystals -- 29. Defects in Cryoelectron Micrographs -- CHAPTER 11. IMAGE RECORDING -- 1. Exposure Time -- 2. Over/Underexposure -- 3. Effects of Development -- 4. Exposure Dose Adjustment -- 5. Primary Magnification -- 6. Damage to Negatives -- 7. Damage to Wet Negatives -- 8. Film/Imaging Plate/Charge-Coupled Device (CCD) Camera -- 9. Enlarged Digital Recordings.

10. Variation in Electron Dose -- 11. Corrections of CCD Camera -- CHAPTER 12. PHOTOGRAPHIC AND DIGITAL PRINTING -- 1. Photographic Paper of Different Grades -- 2. Multigrade Paper -- 3. Exposure and Development -- 4. Enlargement of Micrograph Details -- 5. Objective Lens in Enlarger -- 6. Focusing of Enlarger -- 7. Intermediate Diapositive -- 8. Errors in Photographic Printing -- 9. Retouch -- 10. Comparison of Printers: Low Magnification -- 12. Pixel Size at Printing -- CHAPTER 13. NEGATIVE STAINING -- 1. Negative Staining Methods -- 2. Properties of Support Film -- 3. Comparison of Stains -- 4. Thickness of Stain -- 5. Concentration of Specimen -- 6. Deformation of Specimen -- 7. Radiation Damage -- CHAPTER 14. AUTORADIOGRAPHY -- 1. Undeveloped Emulsion -- 2. Developed Emulsion -- 3. Resolution -- 4. Quantitation -- 5. Preparatory Defects -- CHAPTER 15. CYTOCHEMISTRY -- 1. Influence of Fixation -- 2. Preincubation Treatment -- 3. Appearance of Reaction Product -- 4. Composition of Incubation Medium -- 5. Cytochemical Resolution -- 6. Unspecific Staining -- 7. Extraction of Reaction Product -- CHAPTER 16. IMMUNOCYTOCHEMISTRY -- 1. Fixation of Sensitive Antigens -- 2. Fixation of Insensitive Antigens -- 3. Comparison of Embedding Media -- 4. Influence of Preincubation Solutions -- 5. Comparison of Primary Antibodies -- 6. Dilution of Primary Antibody -- 7. Quantitation of Gold Particles -- 8. Controls -- 9. Comparison of Gold Probes -- 10. Amplification of Gold Particles -- 11. Section Staining -- 12. Resolution -- 13. Background Labeling -- 14. Antigen Retrieval by Etching -- 15. Antigen Retrieval with Sodium Dodecyl Sulfate -- 16. Double Labeling -- 17. Immunonegative Staining -- 18. Freeze-Fracture Replica Labeling -- 19. Preembedding Labeling -- 20. Semithin Light Microscopic Sections -- CHAPTER 17. FREEZE FRACTURING AND SHADOWING.

1. Shadowing of DNA Molecules -- 2. Shadowing of Protein Molecules -- 3. Freeze-Fractured Membrane Faces -- 4. Thickness of Replica: Low Magnification -- 5. Thickness of Replica: High Magnification -- 6. Rotary Shadowing -- 7. Complementary Replicas and Stereo Images -- 8. Ice Crystals and Etching -- 9. Quick-Freeze Deep Etching -- 10. Identification of Transport Molecules -- 11. Contamination -- 12. Plastic Distortion -- 13. Replica Defects -- CHAPTER 18. SAMPLING AND QUANTITATION -- 1. Calibration of Magnification -- 2. Sampling and Object Variability -- 3. Sampling of Pellets: Differential Centrifugation -- 4. Sampling of Pellets: Gradient Centrifugation -- 5. Micrograph Montages -- 6. Automated Digital Montages -- 7. Resolution of Digital Montages -- 8. Measurements on Digital Images -- 9. Stereological Grids -- 10. Cycloid Test System -- CHAPTER 19. IMAGE PROCESSING -- 1. Digital Contrast Changes -- 2. Processing of Scanned Image -- 3. Translational Image Enforcement -- 4. Averaging of Macromolecular Assemblies -- 5. Rotational Image Enforcement -- 6. Photographic versus Computer Averaging -- 7. Fourier Correction of Section Chatter -- 8. Removal of Image Defects -- 9. Scientific Fraud: Removal of Objects -- 10. Scientific Fraud: Manipulation of Labeling -- CHAPTER 20. THREE-DIMENSIONAL RECONSTRUCTIONS -- 1. Comparison between Transmission and Scanning Electron Microscopy -- 2. Serial Sectioning -- 3. Large Three-Dimensional Objects -- 4. Tilting of Section Cell Nucleus -- 5. Tilting of Section" Nuclear Envelope -- 6. Helical Structures -- 7. Computer-Analyzed Helices -- 8. A Three-Dimensional Model of Na, K-ATPase -- APPENDIX: PRACTICAL METHODS -- 1. Fixation -- 2. Dehydration and Embedding -- 3. Low Temperature Embedding -- 4. Support Films -- 5. Ultramicrotomy -- 6. Section Staining -- 7. Microscopy and Image Recording -- 8. Photographic Work.

9. Negative Staining.