This guide provides extensive coverage of microscopic imaging principles. After reviewing the main principles of image formation, diffraction, interference, and polarization used in microscopy, this guide describes the most widely applied microscope configurations and applications. It also covers major system components, including light sources, illumination layouts, microscope optics, and image detection electronics.
This guide also provides a comprehensive overview of microscopy techniques, including bright field and dark field imaging, contrast enhancement methods (such as phase and amplitude contrast), DIC, polarization, and fluorescence microscopy. In addition, it describes scanning techniques (such as confocal and multiphoton imaging points); new trends in super-resolution methods (such as 4Pi microscopy, STED, STORM, and structured illumination); and array microscopy, CARS, and SPIM.
- Glossary of Symbols
- Basic Concepts
- The Nature of Light
- Spectrum of Light vs. Microscopy
- Wave Equations
- Wavefront Propagation
- Optical Path Length (OPL)
- Reflection and Refraction Laws
- Total Internal Reflection
- Evanescent Wave in TIR
- Propagation through Anisotropic Media
- Polarization of Light and Polarization States
- Coherence and Monochromatic Light
- Interference
- Contrast versus Spatial and Temporal Coherence
- Contrast of Fringes (Polarization and Amplitude Ratio)
- Multiple Wave Interference
- Interferometers
- Diffraction
- Diffraction Grating
- Useful Definitions of Geometrical Optics
- Image Formation
- Magnifications
- Stops and Rays in Optical System
- Aberrations
- Chromatic Aberrations
- Spherical and Coma Aberrations
- Astigmatism, Field Curvature and Distortion
- Performance Metrics
- Microscope Construction
- Compound Microscope
- The Eye
- Upright and Inverted Microscopes
- Finite Tube Length Microscope
- Infinity Corrected Systems
- Telecentricity of a Microscope
- Magnification of the Microscope
- Numerical Aperture
- Rayleigh versus Abbe Resolution Limit
- Useful Magnification
- Depth of Field and Depth of Focus
- Magnification and Frequency versus DOF
- Koehler Illumination
- Alignment of Koehler Illumination
- Critical Illumination
- Stereo Microscopes
- Eyepieces
- Nomenclature and Marking of Objectives
- Objective Designs
- Special Objectives and Features
- Special Lens Components
- Cover Glass and Immersion
- Common Light Sources for Microscopy
- LED Light Sources
- Filters
- Polarizers and Polarization Prisms
- Specialized Techniques
- Microscope Objects
- Selection of Microscopy Technique
- Image Comparison
- Phase Contrast
- Visibility in Phase Contrast
- The Phase Contrast Microscope
- Characteristic Features of Phase Contrast Microscopy
- Amplitude Contrast
- Oblique Illumination
- Modulation Contrast - Hoffman Contrast
- Dark Field Microscopy
- Optical Staining: Rheinberg Illumination
- Optical Staining: Dispersion Staining
- Shearing Interferometry - Principle of DIC
- DIC Microscope Design
- Appearance of DIC Images
- Reflectance DIC
- Polarization Microscopy
- Images obtained with Polarization Microscopes
- Compensators
- Confocal Microscopy
- Scanning Approaches
- Images from Confocal Microscope
- Fluorescence
- Configuration of Fluorescence Microscope
- Images from Fluorescence Microscopy
- Properties of Fluorophores
- Single versus Multi-Photon Excitation
- Light Sources for Scanning Microscopy
- Practical Considerations on LSM
- Short Coherence Microscopy
- Optical Profiling Techniques
- Optical Profilometry - System Design
- PSF Algorithms
- Resolution Enhancement Techniques
- Structured Illumination - Axial Sectioning
- Structured Illumination - Resolution Enhancement
- TIRF Microscopy
- Solid Immersion
- Stimulated Emission Depletion
- STORM
- 4Pi Microscopy
- Limits of Light Microscopy
- Other Techniques
- Raman and CARS Microscopy
- SPIM
- Array Microscopy
- Digital Microscopy and CCD
- Digital Microscopy
- CCD Principle
- CCD Architectures
- Noise in CCD
- SNR and Digitization of CCD
- CCD Sampling
- Equation Summary
- Bibliography
- Index
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