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Spie Press Book

Field Guide to Microscopy
Author(s): Tomasz S. Tkaczyk
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Book Description

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.

Book Details

Date Published: 18 May 2010
Pages: 156
ISBN: 9780819472465
Volume: FG13

Table of Contents
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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
Contrast versus Spatial and Temporal Coherence
Contrast of Fringes (Polarization and Amplitude Ratio)
Multiple Wave Interference
Diffraction Grating
Useful Definitions of Geometrical Optics
Image Formation
Stops and Rays in Optical System
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
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
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
Confocal Microscopy
Scanning Approaches
Images from Confocal Microscope
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
4Pi Microscopy
Limits of Light Microscopy
Other Techniques
Raman and CARS Microscopy
Array Microscopy
Digital Microscopy and CCD
Digital Microscopy
CCD Principle
CCD Architectures
Noise in CCD
SNR and Digitization of CCD
CCD Sampling
Equation Summary

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