An Introduction to Microscopy by Means of Light, Electrons, X-Rays, or Ultrasound

Many people look upon a microscope as a mere instrument(l); to them microscopy is instrumentation. Other people consider a microscope to be simply an aid to the eye; to them microscopy is primarily an expan­ sion of macroscopy. In actuality, microscopy is both objective and sub­ jective; it is seeing through an instrument by means of the eye, and more importantly, the brain. The function of the brain is to interpret the eye's image in terms of the object's structure. Thought and experience are required to distinguish structure from artifact. It is said that Galileo (1564-1642) had his associates first look through his telescope­ microscope at very familiar objects to convince them that the image was a true representation of the object. Then he would have them proceed to hitherto unknown worlds too far or too small to be seen with the un­ aided eye. Since Galileo's time, light microscopes have been improved so much that performance is now very close to theoretical limits. Electron microscopes have been developed in the last four decades to exhibit thousands of times the resolving power of the light microscope. Through the news media everyone is made aware of the marvelous microscopical accomplishments in imagery. However, little or no hint is given as to what parts of the image are derived from the specimen itself and what parts are from the instrumentation, to say nothing of the changes made during preparation of the specimen.

1. A Brief History of Microscopy.- 1.1. Introduction.- 1.2. Corrections for Aberrations.- 1.3. Dark-Field Microscopy.- 1.4. Polarizing Microscope.- 1.5. The Short History of Electron Microscopes.- 1.6. Brief History of the Scanning Electron Microscope (SEM).- 1.7. The Electron-Probe Microanalyzer (EPMA).- 1.8. History of Field-Emission Microscopes.- 1.9. Scanning Acoustic Microscopy.- 2. Definitions, Attributes Contributing to Visibility, and Principles.- 2.1. Definitions and Attributes Contributing to Visibility.- 2.2. Principles in Terms of Attributes.- 2.2.1. Thought, Memory, and Imagination.- 2.2.2. Resolving Power.- 2.2.3. Resolution.- 2.2.4. Contrast Perception.- 2.2.5. Refractive Aberrations.- 2.2.6. Cleanliness and Orderliness.- 2.2.7. Depth of Focus.- 2.2.8. Focus.- 2.2.9. Illumination.- 2.2.10. Radiation.- 2.2.11. Anisotropy.- 2.2.12. Magnification.- 2.2.13. Field of View.- 2.2.14. Antiglare Devices.- 2.2.15. Cues to Depth.- 2.2.16. Working Distance.- 2.2.17. Depth of Field.- 2.2.18. Structure of the Specimen.- 2.2.19. Morphology of the Specimen.- 2.2.20. Information about the Specimen.- 2.2.21. Experimentation.- 2.2.22. Preparation of the Specimen.- 2.2.23. Behavior of the Specimen.- 2.2.24. Photomicrography.- 2.3. Summary.- 3. Simple and Compound Microscopes.- 3.1. The Limiting Resolution.- 3.2. Simple Microscope: One Lens System.- 3.3. Compound Microscope: Two or More Lens Systems.- 3.4. Stereo Compound Microscopes.- 3.5. The Biological Microscope.- 3.5.1. Attitudes.- 3.5.2. Experience.- 3.5.3. Learning and Teaching.- 3.5.4. Resolving Power.- 3.5.5. Resolution.- 3.5.6. Contrast.- 3.5.7. Corrections for Aberrations.- 3.5.8. Cleanliness.- 3.5.9. Depth of Focus.- 3.5.10. Focusing.- 3.5.11. Illumination.- 3.5.12. Radiation.- 3.5.13. Anisotropy.- 3.5.14. Magnification.- 3.5.15. Field of View.- 3.5.16. Antiglare.- 3.5.17. Cues to Depth.- 3.5.18. Depth of Field.- 3.5.19. Working Distance.- 3.5.20. Structure of the Specimen.- 3.5.21. Morphology of the Specimen.- 3.5.22. Information about the Specimen.- 3.5.23. Experimentation.- 3.5.24. Preparation of the Specimen.- 3.5.25. Behavior of the Specimen.- 3.5.26. Photomicrography.- 3.6. Summary.- 4. Compound Microscopes Using Reflected Light.- 4.1. Study of Surfaces by Reflected Light.- 4.2. Resolving Power.- 4.3. Contrast.- 4.4. Corrections for Aberrations.- 4.5. Cleanliness of the Specimen.- 4.6. Depth of Focus.- 4.7. Role of Focus.- 4.8. Illumination.- 4.9. Radiation.- 4.10. Magnification.- 4.11. Field of View.- 4.12. Problems of Glare.- 4.13. Cue to Depth.- 4.14. Depth of Field.- 4.15. Working Distance.- 4.16. Study of Structure.- 4.17. Study of Morphology.- 4.18. Information about the Specimen.- 4.19. Experimentation.- 4.20. Behavior of the Specimen.- 4.21. Preparation of the Specimen.- 4.22. Photomicrographic Techniques.- 4.23. Summary.- 5. Microscopy with Polarized Light.- 5.1. The Overhead Projector.- 5.2. Anisotropy.- 5.3. Numerical Aperture and Interference Figures.- 5.4. Resolution: Interaction of Specimen and Polarized Light.- 5.5. Contrast: Michel-Levy Interference Chart.- 5.5.1. Retardation Plates.- 5.5.2. Thickness of the Specimen.- 5.6. Correction for Aberrations Due to Strain.- 5.7. Cleanliness: Freedom from Interference Films.- 5.8. Depth of Focus.- 5.9. Focus.- 5.10. Illumination.- 5.11. Radiation.- 5.12. Magnification.- 5.13. Field of View of an Interference Figure.- 5.14. Glare.- 5.15. Cues to Depth.- 5.16. Depth of Field.- 5.17. Working Distance.- 5.18. Structure of the Specimen.- 5.19. Morphology of the Specimen.- 5.20. Information about the Specimen.- 5.21. Experimentation.- 5.22. Behavior of the Specimen.- 5.23. Preparation of the Specimen.- 5.24. Photography.- 5.25. Summary.- 6. The Microscopical Properties of Fibers.- 6.1. Introduction.- 6.2. The Eight Optical Properties of Fibers.- 6.3. Molecular Anisotropy.- 6.4. Anisotropy: Molecular Orientation and Organization.- 6.5. Transparent Sheets, Foils, and Films.- 6.6. Summary.- 7. Microscopical Properties of Crystals.- 7.1. Structural Classifications.- 7.2. Morphology.- 7.3. Miller Indices.- 7.4. Isomorphism.- 7.5. Skeletal Morphology.- 7.6. Isotropic System.- 7.7. Uniaxial Crystals.- 7.8. Biaxial Crystals.- 7.9. Optical Properties of the Liquid-Crystalline or Mesomorphic State.- 7.10. Thermotropic, Mesomorphic, Single Compounds.- 7.11. Kinds of Morphology (Texture).- 7.11.1. Homeotropic Textures.- 7.11.2. Focal Conic Textures.- 7.11.3. Other Smectic Textures.- 7.11.4. Nematic Textures.- 7.11.5. Cholesteric Textures.- 7.12. Lyotropic Phases.- 7.13. Summary.- 8. Photomicrography.- 8.1. Photomicrograph: Image Produced by Light, Electrons, or X-Rays.- 8.2. Two Attitudes: Artistic versus Scientific.- 8.3. Experience: Records of Negatives.- 8.4. Imagination.- 8.5. Resolving Power.- 8.6. Resolution by Photomacrographic Lenses.- 8.7. Contrast: Nature of Photosensitive Materials.- 8.8. Corrections for Aberrations.- 8.9. Cleanliness in the Darkroom.- 8.10. Depth of Focus.- 8.11. Focusing.- 8.12. Illumination.- 8.13. Spectrum of Effective Radiation.- 8.14. Anisotropy.- 8.15. Useful Magnification in Photomacrographs.- 8.16. Field of View.- 8.17. Summary.- 9. Contrast: Phase, Amplitude, and Color.- 9.1. Contrast: Colorless and Color.- 9.2. Interference: Destructive and Constructive.- 9.3. Phase-Amplitude Contrast.- 9.4. Phase-Amplitude Contrast in Determining Refractive Index.- 9.5. Variable Phase-Amplitude Microscopy.- 9.6. Modulation-Contrast Microscopy.- 9.7. Dispersion Staining.- 9.8. Special Accessories.- 9.9. The Schlieren Microscope.- 9.10. Summary.- 10. Interferometry in Microscopy.- 10.1. Interference of Two Whole Beams.- 10.2. Kinds of Interference Microscopes.- 10.2.1. Single Microscopes.- 10.2.2. Double Microscopes.- 10.3. Applications to Highly Birefringent Specimens.- 10.4. Summary.- 11. Microscopical Stages.- 11.1. Introduction.- 11.2. Micromanipulators.- 11.3. Heatable Stages.- 11.3.1. Hot Stages with Long Working Distances.- 11.3.2. Hot Stages with Short Working Distances.- 11.3.3. Hot-Wire Stage.- 11.4. Very Hot Stage.- 11.5. Cold Stages.- 11.6. Other Special Cells and Cuvettes.- 11.7. Summary.- 12. Transmission Electron Microscopy.- 12.1. Electron Microscopes.- 12.2. Electron Lenses.- 12.3. Resolving Power.- 12.4. Resolution.- 12.5. Contrast.- 12.6. Aberrations.- 12.7. Cleanliness.- 12.8. Depth of Focus.- 12.9. Focus.- 12.10. Illumination.- 12.11. Anisotropy.- 12.12. Useful Magnification.- 12.13. Field of View.- 12.14. Artifacts.- 12.15. Cues to Depth.- 12.16. Thickness of the Specimen.- 12.17. Depth of Field.- 12.18. Structure of the Specimen.- 12.19. Morphology of the Specimen.- 12.20. Information about the Specimen.- 12.21. Experimentation.- 12.22. Preparation of the Specimen.- 12.23. Electron Micrography.- 12.24. Scanning Transmission Electron Microscope (STEM).- 12.25. Summary.- 13. Scanning Electron Microscopy.- 13.1. Introduction.- 13.2. Resolving Power.- 13.3. Resolution.- 13.4. Contrast.- 13.5. Aberrations.- 13.6. Cleanliness.- 13.7. Depth of Focus.- 13.8. Focusing.- 13.9. Illumination.- 13.10. Radiation.- 13.11. Useful Magnification.- 13.12. Field of View.- 13.13. Noise.- 13.14. Cues to Depth.- 13.15. Working Distance.- 13.16. Depth of Field.- 13.17. Structure.- 13.18. Morphology.- 13.19. Information.- 13.20. Dynamic Experimentation.- 13.21. Behavior of the Specimen.- 13.22. Preparation of the Specimen.- 13.23. Photomicrography.- 13.24. Summary.- 14. Field-Emission Microscopes.- 14.1. Introduction.- 14.2. Attributes Contributing to Visibility by Field-Emission Microscopy.- 14.2.1. Thought, Memory, and Imagination.- 14.2.2. Resolving Power.- 14.2.3. Resolution.- 14.2.4. Contrast.- 14.2.5. Aberrations.- 14.2.6. Cleanliness.- 14.2.7. Depth of Focus.- 14.2.8. Illumination.- 14.2.9. Radiation.- 14.2.10. Magnification.- 14.2.11. Field of View.- 14.2.12. Artifacts.- 14.2.13. Working Distance.- 14.2.14. Depth of Field.- 14.2.15. Structure.- 14.2.16. Morphology.- 14.2.17. Information.- 14.2.18. Experiments.- 14.2.19. Behavior.- 14.2.20. Preparation of the Specimen.- 14.3. Summary.- 15. X-Ray Microscopy.- 15.1. X-Rays.- 15.2. Condenser Lenses.- 15.3. X-Ray Holography.- 15.4. Summary.- 16. Acoustic Microscopy.- 16.1. Ultrasound Waves from Microspecimens.- 16.2. Acoustic Microscopes: SLAM versus SAM.- 16.2.1. Theoretical Resolving Power of the Acoustic Microscope.- 16.2.2. Practical Resolution of the Acoustic Microscope.- 16.2.3. Contrast in Acoustic Images.- 16.2.4. Aplanatic Lenses in SAM.- 16.2.5. Cleanliness in Acoustic Microscopes.- 16.2.6. Depth of Focus in SAM.- 16.2.7. Focusing SAM.- 16.2.8. Acoustic Radiation.- 16.2.9. Magnification.- 16.2.10. Field of View.- 16.2.11. Stray Acoustic Radiation.- 16.2.12. Three-Dimensional Aspect of SLAM.- 16.2.13. Thickness of the Specimen.- 16.2.14. Working Distance.- 16.2.15. Structure of the Specimen.- 16.2.16. Anisotropy.- 16.2.17. Morphology of the Specimen.- 16.2.18. Information about the Acoustical Image.- 16.2.19. Experiments with the Specimen.- 16.2.20. Behavior of the Specimen.- 16.2.21. Preparation of the Specimen.- 16.2.22. Photomicrography.- 16.3. Summary.- References.- Author Index.