Glossary of Symbols ............................................ xi
Basics Concepts ................................................. 1
Nature of Light .............................................. 1
The Spectrum of Microscopy ................................... 2
Wave Equations ............................................... 3
Wavefront Propagation ........................................ 4
Optical Path Length (OPL) .................................... 5
Laws of Reflection and Refraction ............................ 6
Total Internal Reflection .................................... 7
Evanescent Wave in Total Internal Reflection ................. 8
Propagation of Light in Anisotropic Media .................... 9
Polarization of Light and Polarization States ............... 10
Coherence and Monochromatic Light ........................... 11
Interference ................................................ 12
Contrast vs Spatial and Temporal Coherence .................. 13
Contrast of Fringes (Polarization and Amplitude Ratio) ... 15
Multiple Wave Interference .................................. 16
Interferometers ............................................. 17
Diffraction ................................................. 18
Diffraction Grating ......................................... 19
Useful Definitions from Geometrical Optics .................. 21
Image Formation ............................................. 22
Magnification ............................................... 23
Stops and Rays in an Optical System ......................... 24
Aberrations ................................................. 25
Chromatic Aberrations ....................................... 26
Spherical Aberration and Coma ............................... 27
Astigmatism, Field Curvature, and Distortion ................ 28
Performance Metrics ......................................... 29
Microscope Construction ........................................ 31
The Compound Microscope ..................................... 31
The Eye ..................................................... 32
Upright and Inverted Microscopes ............................ 33
The Finite Tube Length Microscope ........................... 34
Infinity-Corrected Systems .................................. 35
Telecentricity of a Microscope .............................. 36
Magnification of a Microscope ............................... 37
Numerical Aperture .......................................... 38
Resolution Limit ............................................ 39
Useful Magnification ........................................ 40
Depth of Field and Depth of Focus ........................... 41
Magnification and Frequency vs Depth of Field ............... 42
Köhler Illumination ......................................... 43
Alignment of Kцhler Illumination ............................ 45
Critical Illumination ....................................... 46
Stereo Microscopes .......................................... 47
Eyepieces ................................................... 48
Nomenclature and Marking of Objectives ...................... 50
Objective Designs ........................................... 51
Special Objectives and Features ............................. 53
Special Lens Components ..................................... 55
Cover Glass and Immersion ................................... 56
Common Light Sources for Microscopy ......................... 58
LED Light Sources ........................................... 59
Filters ..................................................... 60
Polarizers and Polarization Prisms ............................. 61
Specialized Techniques ...................................... 63
Amplitude and Phase Objects ................................. 63
The Selection of a Microscopy Technique ..................... 64
Image Comparison ............................................ 65
Phase Contrast .............................................. 66
Visibility in Phase Contrast ................................ 69
The Phase Contrast Microscope ............................... 70
Characteristic Features of Phase Contrast ................... 71
Amplitude Contrast .......................................... 72
Oblique Illumination ........................................ 73
Modulation Contrast ......................................... 74
Hoffman Contrast ............................................ 75
Dark Field Microscopy ....................................... 76
Optical Staining: Rheinberg Illumination .................... 77
Optical Staining: Dispersion Staining ....................... 78
Shearing Interferometry: The Basis for DIC .................. 79
DIC Microscope Design ....................................... 80
Appearance of DIC Images .................................... 81
Reflectance DIC ............................................. 82
Polarization Microscopy ..................................... 83
Images Obtained with Polarization Microscopes ............... 84
Compensators ................................................ 85
Confocal Microscopy ......................................... 86
Scanning Approaches ......................................... 87
Images from a Confocal Microscope ........................... 89
Fluorescence ................................................ 90
Configuration of a Fluorescence Microscope .................. 91
Images from Fluorescence Microscopy ......................... 93
Properties of Fluorophores .................................. 94
Single vs Multi-Photon Excitation ........................... 95
Light Sources for Scanning Microscopy ....................... 96
Practical Considerations in LSM ............................. 97
Interference Microscopy ..................................... 98
Optical Coherence Tomography/Microscopy ..................... 99
Optical Profiling Techniques ............................... 100
Optical Profilometry: System Design ........................ 101
Phase-Shifting Algorithms .................................. 102
Resolution Enhancement Techniques ............................. 103
Structured Illumination: Axial Sectioning .................. 103
Structured Illumination: Resolution Enhancement ............ 104
TIRF Microscopy ............................................ 105
Solid Immersion ............................................ 106
Stimulated Emission Depletion .............................. 107
STORM ...................................................... 108
4Pi Microscopy ............................................. 109
The Limits of Light Microscopy ................................ 110
Other Special Techniques ................................... 111
Raman and CARS Microscopy .................................. 111
SPIM ....................................................... 112
Array Microscopy .............................................. 113
Digital Microscopy and CCD Detectors ....................... 114
Digital Microscopy ......................................... 114
Principles of CCD Operation ................................ 115
CCD Architectures .......................................... 116
CCD Noise .................................................. 118
Signal-to-Noise Ratio and the Digitization of CCD .......... 119
CCD Sampling .................................................. 120
Equation Summary .............................................. 122
Bibliography .................................................. 128
Index ......................................................... 133
|
