Light and Video Microscopy,
Edition 2Editors: By Randy O. Wayne
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The purpose of this book is to provide the most comprehensive, easy-to-use, and informative guide on light microscopy. Light and Video Microscopy will prepare the reader for the accurate interpretation of an image and understanding of the living cell. With the presentation of geometrical optics, it will assist the reader in understanding image formation and light movement within the microscope. It also provides an explanation of the basic modes of light microscopy and the components of modern electronic imaging systems and guides the reader in determining the physicochemical information of living and developing cells, which influence interpretation.
Key Features
- Brings together mathematics, physics, and biology to provide a broad and deep understanding of the light microscope
- Clearly develops all ideas from historical and logical foundations
- Laboratory exercises included to assist the reader with practical applications
- Microscope discussions include: bright field microscope, dark field microscope, oblique illumination, phase-contrast microscope, photomicrography, fluorescence microscope, polarization microscope, interference microscope, differential interference microscope, and modulation contrast microscope
About the author
By Randy O. Wayne, Plant cell biologist at Cornell University
Dedication
Preface to the Second Edition
Preface to the First Edition
Chapter 1. The Relation Between the Object and the Image
Luminous and Nonluminous Objects
Object and Image
Theories of Vision
Light Travels in Straight Lines
Images Formed in a Camera Obscura: Geometric Considerations
Where Does Light Come From?
How Can the Amount of Light Be Measured?
Web Resources
Chapter 2. The Geometric Relationship Between Object and Image
Reflection by a Plane Mirror
Reflection by a Curved Mirror
Reflection from Various Sources
Images Formed by Refraction at a Plane Surface
Images Formed by Refraction at a Curved Surface
Fermat’s Principle
Optical Path Length
Lens Aberrations
Geometric Optics and Biology
Geometric Optics of the Human Eye
Web Resources
Chapter 3. The Dependence of Image Formation on the Nature of Light
Christiaan Huygens and the Invention of the Wave Theory of Light
Thomas Young and the Development of the Wave Theory of Light
James Clerk Maxwell and the Wave Theory of Light
Ernst Abbe and the Relationship of Diffraction to Image Formation
Resolving Power and the Limit of Resolution
Contrast
Web Resources
Chapter 4. Bright-Field Microscopy
Components of the Microscope
The Optical Paths of the Light Microscope
Using the Bright-Field Microscope
Depth of Field
Out-of-Focus Contrast
Uses of Bright-Field Microscopy
Care and Cleaning of the Light Microscope
Web Resources
Chapter 5. Photomicrography
Setting up the Microscope for Photomicrography
Scientific History of Photography
General Nature of the Photographic Process
The Resolution of the Film
Exposure and Composition
The Similarities between Film and the Retina
Web Resources
Chapter 6. Methods of Generating Contrast
Dark-Field Microscopy
Rheinberg Illumination
Oblique Illumination
Phase-Contrast Microscopy
Oblique Illumination Reconsidered
Annular Illumination
Chapter 7. Polarization Microscopy
What is Polarized Light?
Use an Analyzer to Test for Polarized Light
Production of Polarized Light
Influencing Light
Design of a Polarizing Microscope
What is the Molecular Basis of Birefringence?
Interference of Polarized Light
The Origin of Colors in Birefringent Specimens
Use of Compensators to Determine the Magnitude and Sign of Birefringence
Crystalline versus Form Birefringence
Orthoscopic versus Conoscopic Observations
Reflected Light Polarization Microscopy
Uses of Polarization Microscopy
Optical Rotatory (or Rotary) Polarization and Optical Rotatory (or Rotary) Dispersion
Web Resources
Chapter 8. Interference Microscopy
Generation of Interference Colors
The Relationship of Interference Microscopy to Phase-Contrast Microscopy
Quantitative Interference Microscopy: Determination of the Refractive Index, Mass, Concentration of Dry Matter, Concentration of Water, and Density
Source of Errors When Using an Interference Microscope
Making a Coherent Reference Beam
Double-Beam versus Multiple-Beam Interference
Interference Microscopes Based on a Mach–Zehnder Type Interferometer
Interference Microscopes Based on Polarized Light
The Use of Transmission Interference Microscopy in Biology
Reflection–Interference Microscopy
Uses of Reflection–Interference Microscopy in Biology
Web Resources
Chapter 9. Differential Interference Contrast (DIC) Microscopy
Design of a Transmitted Light Differential Interference Contrast Microscope
Interpretation of a Transmitted Light Differential Interference Contrast Image
Design of a Reflected Light Differential Interference Contrast Microscope
Interpretation of a Reflected Light Differential Interference Contrast Image
Chapter 10. Amplitude Modulation Contrast Microscopy
Hoffman Modulation Contrast Microscopy
Reflected Light Hoffman Modulation Contrast Microscopy
The Single-Sideband Edge Enhancement Microscope
Web Resources
Chapter 11. Fluorescence Microscopy
Discovery of Fluorescence
Physics of Fluorescence
Design of a Fluorescence Microscope
Fluorescence Probes
Pitfalls and Cures in Fluorescence Microscopy
Web Resources
Chapter 12. Various Types of Microscopes and Accessories
Confocal Microscopes
Total Internal Reflectance Fluorescence Microscopy
Superresolution With Wide-Field Microscopes
Fluorescence Resonance Energy Transfer
Fluorescence Redistribution After Photobleaching
Laser Microbeam Microscope
Optical Tweezers
Laser Capture Microdissection
Laser Doppler Microscope
Centrifuge Microscope
X-Ray Microscope
Infrared Microscope
Nuclear Magnetic Resonance Imaging Microscope
Stereo Microscopes
Scanning Probe Microscopes
Acoustic Microscope
Horizontal and Traveling Microscopes
Microscopes for Children
Microscope Accessories
Web Resources
Chapter 13. Video and Digital Microscopy
The Value of Video and Digital Microscopy
Video and Digital Cameras: The Optical-to-Electrical Signal Converters
Monitors: Conversion of an Electronic Signal into an Optical Signal
Storage of Video and Digital Images
Connecting a Video System
Web Resources
Chapter 14. Image Processing and Analysis
Analog Image Processing
Digital Image Processing
Enhancement Functions of Digital Image Processors
Analysis Functions of Digital Image Processors
Toupview Digital Image Processing Software
The Ethics of Digital Image Processing
Web Resources
Chapter 15. Laboratory Exercises
Laboratory 1: The Nature of Light and Geometric Optics
Laboratory 2: Physical Optics
Laboratory 3: The Bright-Field Microscope and Image Formation
Laboratory 4: Phase-Contrast Microscopy, Dark-Field Microscopy, Rheinberg Illumination, and Oblique Illumination
Laboratory 5: Fluorescence Microscopy
Laboratory 6: Polarized Light
Laboratory 7: Polarizing Light Microscopy
Laboratory 8: Interference Microscopy
Laboratory 9: Differential Interference Contrast Microscopy and Hoffman Modulation Contrast Microscopy
Laboratory 10: Video and Digital Microscopy and Analog and Digital Image Processing
Web Resources
References
Appendix I. Light Microscopy: A Retrospective
Web Resource
Appendix II. A Final Exam
Appendix III. A Microscopist’s Model of the Photon
Index
Color Plates
Title Reviews
"As such, this book would be an excellent undergraduate-level text on optical microscopy for biologists. It is also valuable to anyone using a light microscope."—Carol Bayles, Cornell University, Ithaca, NY, USA
